The Experts below are selected from a list of 519 Experts worldwide ranked by ideXlab platform
Ignacio Rodríguez-crespo - One of the best experts on this subject based on the ideXlab platform.
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LC8 dynein light chain (DYNLL1) binds to the C-terminal domain of ATM-interacting protein (ATMIN/ASCIZ) and regulates its subcellular localization.
Biochemical and biophysical research communications, 2011Co-Authors: Péter Rapali, María Flor García-mayoral, Krisztian Tarnok, Katalin Schlett, Juan Pablo Albar, Marta Bruix, Laszlo Nyitray, Mónica Martínez-moreno, Ignacio Rodríguez-crespoAbstract:Highlights: Black-Right-Pointing-Pointer We have screened a human library with dynein light chain DYNLL1 (DLC8) as bait. Black-Right-Pointing-Pointer Dynein light chain DYNLL1 binds to ATM-kinase interacting protein (ATMIN). Black-Right-Pointing-Pointer ATMIN has 17 SQ/TQ motifs, a motif frequently found in DYNLL1-binding partners. Black-Right-Pointing-Pointer The two proteins interact in vitro, with ATMIN displaying at least five binding sites. Black-Right-Pointing-Pointer The interaction of ATMIN and DYNNL1 in transfected cells can also be observed. -- Abstract: LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of themore » DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.« less
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The association of viral proteins with host cell dynein components during virus infection
FEBS Journal, 2011Co-Authors: Javier Merino-gracia, María Flor García-mayoral, Ignacio Rodríguez-crespoAbstract:After fusion with the cellular plasma membrane or endosomal membranes, viral particles are generally too large to diffuse freely within the crowded cytoplasm environment. Thus, they will never reach the cell nucleus or the perinuclear areas where replication or reverse transcription usually takes place. It has been proposed that many unrelated viruses are transported along microtubules in a retrograde manner using the cellular dynein machinery or, at least, some dynein components. A putative employment of the dynein motor in a dynein-mediated transport has been suggested from experiments in which viral capsid proteins were used as bait in yeast two-hybrid screens using libraries composed of cellular proteins and dynein-associated chains were retrieved as virus-interacting proteins. In most cases DYNLL1, DYNLT1 or DYNLRB1 were identified as the dynein chains that interact with viral proteins. The importance of these dynein–virus interactions has been supported, in principle, by the observation that in some cases the dynein-interacting motifs of viral proteins altered by site-directed mutagenesis result in non-infective virions. Furthermore, overexpression of p50 dynamitin, which blocks the dynein–dynactin interaction, or incubation of infected cells with peptides that compete with viral polypeptides for dynein binding have been shown to alter the viral retrograde transport. Still, it remains to be proved that dynein light chains can bind simultaneously to incoming virions and to the dynein motor for retrograde transport to take place. In this review, we will analyse the association of viral proteins with dynein polypeptides and its implications for viral infection.
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Structural models of DYNLL1 with interacting partners: African swine fever virus protein p54 and postsynaptic scaffolding protein gephyrin
FEBS letters, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Marta BruixAbstract:DYNLL1, the smallest dynein light chain, interacts with different cargos facilitating their cellular transport. Usually the sequence recognized in the targets is homologous to the GIQVD or the KXTQT motifs with a glutamine that is important for binding. Here we add two new examples of DYNLL1 targets that can be classified into these two groups: ASFV p54 and gephyrin. Using NMR we demonstrate the direct interaction between DYNLL1 and two peptides derived from their interacting sequences. We model the structure of both complexes and show that the overall binding mode is preserved as in other complexes despite differences at the residue-specific interactions.
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Structural basis for the interaction between dynein light chain 1 and the glutamate channel homolog GRINL1A.
The FEBS journal, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Juan Pablo Albar, Mónica Martínez-moreno, Marta BruixAbstract:Human dynein light chain 1 (DYNLL1) is a dimeric 89-residue protein that is known to be involved in cargo binding within the dynein multiprotein complex. Over 20 protein targets, of both cellular and viral origin, have been shown to interact with DYNLL1, and some of them are transported in a retrograde manner along microtubules. Using DYNLL1 as bait in a yeast two-hybrid screen with a human heart library, we identified GRINL1A (ionotropic glutamate receptor N-methyl-d-aspartate-like 1A), a homolog of the ionotropic glutamate receptor N-methyl d-aspartate, as a DYNLL1 binding partner. Binding of DYNLL1 to GRINL1A was also demonstrated using GST fusion proteins and pepscan membranes. Progressive deletions allowed us to narrow the DYNLL1 binding region of GRINL1A to the sequence REIGVGCDL. Combining these results with NMR data, we have modelled the structure of the GRINL1A–DYNLL1 complex. By analogy with known structures of DYNLL1 bound to BCL-2-interacting mediator (BIM) or neuronal nitric oxide synthase (nNOS), the GRINL1A peptide also adopts an extended β-strand conformation that expands the central β-sheet within DYNLL1. Structural comparison with the nNOS–DYNLL1 complex reveals that a glycine residue of GRINL1A occupies the conserved glutamine site within the DYNLL1 binding groove. Hence, our data identify a novel membrane-associated DYNLL1 binding partner and suggest that additional DYNLL1-binding partners are present near this glutamate channel homolog. Structured digital abstract • MINT-7713396: DYNLL1 (uniprotkb:P63167) and GRINL1A (uniprotkb:P0CAP1) bind (MI:0407)by nuclear magnetic resonance (MI:0077) • MINT-7713280, MINT-7713382: DYNLL1, (uniprotkb:P63167) physically interacts (MI:0915) with GRINL1A (uniprotkb:P0CAP1) by two hybrid (MI:0018) • MINT-7713416, MINT-7713439: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by peptide array (MI:0081) • MINT-7713307: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by pull down (MI:0096)
Marta Bruix - One of the best experts on this subject based on the ideXlab platform.
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lc8 dynein light chain DYNLL1 binds to the c terminal domain of atm interacting protein atmin asciz and regulates its subcellular localization
Biochemical and Biophysical Research Communications, 2011Co-Authors: Péter Rapali, Maria Flor Garciamayoral, Monica Martinezmoreno, Krisztian Tarnok, Katalin Schlett, Juan Pablo Albar, Marta Bruix, Laszlo Nyitray, Ignacio RodriguezcrespoAbstract:Highlights: Black-Right-Pointing-Pointer We have screened a human library with dynein light chain DYNLL1 (DLC8) as bait. Black-Right-Pointing-Pointer Dynein light chain DYNLL1 binds to ATM-kinase interacting protein (ATMIN). Black-Right-Pointing-Pointer ATMIN has 17 SQ/TQ motifs, a motif frequently found in DYNLL1-binding partners. Black-Right-Pointing-Pointer The two proteins interact in vitro, with ATMIN displaying at least five binding sites. Black-Right-Pointing-Pointer The interaction of ATMIN and DYNNL1 in transfected cells can also be observed. -- Abstract: LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of themore » DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.« less
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lc8 dynein light chain DYNLL1 binds to the c terminal domain of atm interacting protein atmin asciz and regulates its subcellular localization
Biochemical and Biophysical Research Communications, 2011Co-Authors: Péter Rapali, Maria Flor Garciamayoral, Monica Martinezmoreno, Krisztian Tarnok, Katalin Schlett, Juan Pablo Albar, Marta Bruix, Laszlo Nyitray, Ignacio RodriguezcrespoAbstract:Abstract LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of the DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.
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LC8 dynein light chain (DYNLL1) binds to the C-terminal domain of ATM-interacting protein (ATMIN/ASCIZ) and regulates its subcellular localization.
Biochemical and biophysical research communications, 2011Co-Authors: Péter Rapali, María Flor García-mayoral, Krisztian Tarnok, Katalin Schlett, Juan Pablo Albar, Marta Bruix, Laszlo Nyitray, Mónica Martínez-moreno, Ignacio Rodríguez-crespoAbstract:Highlights: Black-Right-Pointing-Pointer We have screened a human library with dynein light chain DYNLL1 (DLC8) as bait. Black-Right-Pointing-Pointer Dynein light chain DYNLL1 binds to ATM-kinase interacting protein (ATMIN). Black-Right-Pointing-Pointer ATMIN has 17 SQ/TQ motifs, a motif frequently found in DYNLL1-binding partners. Black-Right-Pointing-Pointer The two proteins interact in vitro, with ATMIN displaying at least five binding sites. Black-Right-Pointing-Pointer The interaction of ATMIN and DYNNL1 in transfected cells can also be observed. -- Abstract: LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of themore » DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.« less
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Structural models of DYNLL1 with interacting partners: African swine fever virus protein p54 and postsynaptic scaffolding protein gephyrin
FEBS letters, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Marta BruixAbstract:DYNLL1, the smallest dynein light chain, interacts with different cargos facilitating their cellular transport. Usually the sequence recognized in the targets is homologous to the GIQVD or the KXTQT motifs with a glutamine that is important for binding. Here we add two new examples of DYNLL1 targets that can be classified into these two groups: ASFV p54 and gephyrin. Using NMR we demonstrate the direct interaction between DYNLL1 and two peptides derived from their interacting sequences. We model the structure of both complexes and show that the overall binding mode is preserved as in other complexes despite differences at the residue-specific interactions.
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Structural basis for the interaction between dynein light chain 1 and the glutamate channel homolog GRINL1A.
The FEBS journal, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Juan Pablo Albar, Mónica Martínez-moreno, Marta BruixAbstract:Human dynein light chain 1 (DYNLL1) is a dimeric 89-residue protein that is known to be involved in cargo binding within the dynein multiprotein complex. Over 20 protein targets, of both cellular and viral origin, have been shown to interact with DYNLL1, and some of them are transported in a retrograde manner along microtubules. Using DYNLL1 as bait in a yeast two-hybrid screen with a human heart library, we identified GRINL1A (ionotropic glutamate receptor N-methyl-d-aspartate-like 1A), a homolog of the ionotropic glutamate receptor N-methyl d-aspartate, as a DYNLL1 binding partner. Binding of DYNLL1 to GRINL1A was also demonstrated using GST fusion proteins and pepscan membranes. Progressive deletions allowed us to narrow the DYNLL1 binding region of GRINL1A to the sequence REIGVGCDL. Combining these results with NMR data, we have modelled the structure of the GRINL1A–DYNLL1 complex. By analogy with known structures of DYNLL1 bound to BCL-2-interacting mediator (BIM) or neuronal nitric oxide synthase (nNOS), the GRINL1A peptide also adopts an extended β-strand conformation that expands the central β-sheet within DYNLL1. Structural comparison with the nNOS–DYNLL1 complex reveals that a glycine residue of GRINL1A occupies the conserved glutamine site within the DYNLL1 binding groove. Hence, our data identify a novel membrane-associated DYNLL1 binding partner and suggest that additional DYNLL1-binding partners are present near this glutamate channel homolog. Structured digital abstract • MINT-7713396: DYNLL1 (uniprotkb:P63167) and GRINL1A (uniprotkb:P0CAP1) bind (MI:0407)by nuclear magnetic resonance (MI:0077) • MINT-7713280, MINT-7713382: DYNLL1, (uniprotkb:P63167) physically interacts (MI:0915) with GRINL1A (uniprotkb:P0CAP1) by two hybrid (MI:0018) • MINT-7713416, MINT-7713439: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by peptide array (MI:0081) • MINT-7713307: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by pull down (MI:0096)
María Flor García-mayoral - One of the best experts on this subject based on the ideXlab platform.
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LC8 dynein light chain (DYNLL1) binds to the C-terminal domain of ATM-interacting protein (ATMIN/ASCIZ) and regulates its subcellular localization.
Biochemical and biophysical research communications, 2011Co-Authors: Péter Rapali, María Flor García-mayoral, Krisztian Tarnok, Katalin Schlett, Juan Pablo Albar, Marta Bruix, Laszlo Nyitray, Mónica Martínez-moreno, Ignacio Rodríguez-crespoAbstract:Highlights: Black-Right-Pointing-Pointer We have screened a human library with dynein light chain DYNLL1 (DLC8) as bait. Black-Right-Pointing-Pointer Dynein light chain DYNLL1 binds to ATM-kinase interacting protein (ATMIN). Black-Right-Pointing-Pointer ATMIN has 17 SQ/TQ motifs, a motif frequently found in DYNLL1-binding partners. Black-Right-Pointing-Pointer The two proteins interact in vitro, with ATMIN displaying at least five binding sites. Black-Right-Pointing-Pointer The interaction of ATMIN and DYNNL1 in transfected cells can also be observed. -- Abstract: LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of themore » DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.« less
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The association of viral proteins with host cell dynein components during virus infection
FEBS Journal, 2011Co-Authors: Javier Merino-gracia, María Flor García-mayoral, Ignacio Rodríguez-crespoAbstract:After fusion with the cellular plasma membrane or endosomal membranes, viral particles are generally too large to diffuse freely within the crowded cytoplasm environment. Thus, they will never reach the cell nucleus or the perinuclear areas where replication or reverse transcription usually takes place. It has been proposed that many unrelated viruses are transported along microtubules in a retrograde manner using the cellular dynein machinery or, at least, some dynein components. A putative employment of the dynein motor in a dynein-mediated transport has been suggested from experiments in which viral capsid proteins were used as bait in yeast two-hybrid screens using libraries composed of cellular proteins and dynein-associated chains were retrieved as virus-interacting proteins. In most cases DYNLL1, DYNLT1 or DYNLRB1 were identified as the dynein chains that interact with viral proteins. The importance of these dynein–virus interactions has been supported, in principle, by the observation that in some cases the dynein-interacting motifs of viral proteins altered by site-directed mutagenesis result in non-infective virions. Furthermore, overexpression of p50 dynamitin, which blocks the dynein–dynactin interaction, or incubation of infected cells with peptides that compete with viral polypeptides for dynein binding have been shown to alter the viral retrograde transport. Still, it remains to be proved that dynein light chains can bind simultaneously to incoming virions and to the dynein motor for retrograde transport to take place. In this review, we will analyse the association of viral proteins with dynein polypeptides and its implications for viral infection.
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Structural models of DYNLL1 with interacting partners: African swine fever virus protein p54 and postsynaptic scaffolding protein gephyrin
FEBS letters, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Marta BruixAbstract:DYNLL1, the smallest dynein light chain, interacts with different cargos facilitating their cellular transport. Usually the sequence recognized in the targets is homologous to the GIQVD or the KXTQT motifs with a glutamine that is important for binding. Here we add two new examples of DYNLL1 targets that can be classified into these two groups: ASFV p54 and gephyrin. Using NMR we demonstrate the direct interaction between DYNLL1 and two peptides derived from their interacting sequences. We model the structure of both complexes and show that the overall binding mode is preserved as in other complexes despite differences at the residue-specific interactions.
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Structural basis for the interaction between dynein light chain 1 and the glutamate channel homolog GRINL1A.
The FEBS journal, 2010Co-Authors: María Flor García-mayoral, Ignacio Rodríguez-crespo, Juan Pablo Albar, Mónica Martínez-moreno, Marta BruixAbstract:Human dynein light chain 1 (DYNLL1) is a dimeric 89-residue protein that is known to be involved in cargo binding within the dynein multiprotein complex. Over 20 protein targets, of both cellular and viral origin, have been shown to interact with DYNLL1, and some of them are transported in a retrograde manner along microtubules. Using DYNLL1 as bait in a yeast two-hybrid screen with a human heart library, we identified GRINL1A (ionotropic glutamate receptor N-methyl-d-aspartate-like 1A), a homolog of the ionotropic glutamate receptor N-methyl d-aspartate, as a DYNLL1 binding partner. Binding of DYNLL1 to GRINL1A was also demonstrated using GST fusion proteins and pepscan membranes. Progressive deletions allowed us to narrow the DYNLL1 binding region of GRINL1A to the sequence REIGVGCDL. Combining these results with NMR data, we have modelled the structure of the GRINL1A–DYNLL1 complex. By analogy with known structures of DYNLL1 bound to BCL-2-interacting mediator (BIM) or neuronal nitric oxide synthase (nNOS), the GRINL1A peptide also adopts an extended β-strand conformation that expands the central β-sheet within DYNLL1. Structural comparison with the nNOS–DYNLL1 complex reveals that a glycine residue of GRINL1A occupies the conserved glutamine site within the DYNLL1 binding groove. Hence, our data identify a novel membrane-associated DYNLL1 binding partner and suggest that additional DYNLL1-binding partners are present near this glutamate channel homolog. Structured digital abstract • MINT-7713396: DYNLL1 (uniprotkb:P63167) and GRINL1A (uniprotkb:P0CAP1) bind (MI:0407)by nuclear magnetic resonance (MI:0077) • MINT-7713280, MINT-7713382: DYNLL1, (uniprotkb:P63167) physically interacts (MI:0915) with GRINL1A (uniprotkb:P0CAP1) by two hybrid (MI:0018) • MINT-7713416, MINT-7713439: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by peptide array (MI:0081) • MINT-7713307: GRINL1A (uniprotkb:P0CAP1) binds (MI:0407) to DYNLL1 (uniprotkb:P63167) by pull down (MI:0096)
Silvia E. Racedo - One of the best experts on this subject based on the ideXlab platform.
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Effects of follicle size and stages of maturation on mRNA expression in bovine in vitro matured oocytes.
Molecular Reproduction and Development, 2007Co-Authors: Silvia E. Racedo, Doris Herrmann, Daniel F. Salamone, Christine Wrenzycki, Heinrich NiemannAbstract:Transcription in bovine oocytes: The goal of this study was to unravel the dynamics of transcripts thought to be critically involved in oocyte maturation. The relative abundance (RA) of DYNLL1 (cytoplasmic dynein light chain LC8), DYNC1I1 (cytoplasmic dynein 1 intermediate chain), DCTN1 (dynactin 1; pGlued homolog, the activator of the cytoplasmic dynein complex 1), PMSB1 (proteasome beta subunit 1), PMSA4 (proteasome alfa subunit 4), PAP (poly-A polymerase) and Cx43 (connexin 43) were determined by semi-quantitative endpoint RTPCR at different stages of lVM, that is, GV, GVBD, MI and Mll in oocytes collected from follicles of two different size categories, that is,
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352 EFFECTS OF FOLLICLE SIZE AND STAGE OF MATURATION ON mRNA EXPRESSION IN BOVINE IN VITRO-MATURED OOCYTES
Reproduction Fertility and Development, 2007Co-Authors: Silvia E. Racedo, Doris Herrmann, Daniel F. Salamone, Christine Wrenzycki, Heiner NiemannAbstract:A well-orchestrated expression of genes is required to ensure mammalian oocyte progression from the first meiotic arrest to the metaphase II stage to achieve full developmental competence. DYNLL1 (cytoplasmic dynein light chain LC8), PMSB1 (proteasome beta subunit 1), and DYNC1I1 (cytoplasmic dynein 1 intermediate chain) are crucial genes for nuclear and cytoplasmic maturation. DYNLL1 and DYNC1I1 are constituents of the cytoplasmic dynein 1 complex, the main transport system of the cell. PMSB1 is a subunit of proteasome 20S that is the catalytic core of 26S proteasomes and belongs to the ubiquitin-dependent proteolytic system required for protein degradation. The goal of this study was to compare the relative abundance (RA) of the transcripts of the above genes in oocytes collected from different size follicles at different stages of IVM: germinal vesicle (GV), GV breakdown (GVBD), MI, and MII. Ovaries were collected at a local abattoir. Cumulus–oocyte complexes (COCs) were aspirated from follicles either
Joerg Heierhorst - One of the best experts on this subject based on the ideXlab platform.
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The Novel Zinc Finger Protein dASCIZ Regulates Mitosis in Drosophila via an Essential Role in Dynein Light-Chain Expression
Genetics, 2013Co-Authors: Olga Zaytseva, Nora Tenis, Naomi C Mitchell, Shin-ichiro Kanno, Akira Yasui, Joerg Heierhorst, Leonie M. QuinnAbstract:The essential zinc finger protein ASCIZ (also known as ATMIN, ZNF822) plays critical roles during lung organogenesis and B cell development in mice, where it regulates the expression of dynein light chain (DYNLL1/LC8), but its functions in other species including invertebrates are largely unknown. Here we report the identification of the Drosophila ortholog of ASCIZ (dASCIZ) and show that loss of dASCIZ function leads to pronounced mitotic delays with centrosome and spindle positioning defects during development, reminiscent of impaired dynein motor functions. Interestingly, similar mitotic and developmental defects were observed upon knockdown of the DYNLL/LC8-type dynein light chain Cutup (Ctp), and dASCIZ loss-of-function phenotypes could be suppressed by ectopic Ctp expression. Consistent with a genetic function of dASCIZ upstream of Ctp, we show that loss of dASCIZ led to reduced endogenous Ctp mRNA and protein levels and dramatically reduced Ctp–LacZ reporter gene activity in vivo, indicating that dASCIZ regulates development and mitosis as a Ctp transcription factor. We speculate that the more severe mitotic defects in the absence of ASCIZ in flies compared to mice may be due to redundancy with a second, ASCIZ-independent, Dynll2 gene in mammals in contrast to a single Ctp gene in Drosophila. Altogether, our data demonstrate that ASCIZ is an evolutionary highly conserved transcriptional regulator of dynein light-chain levels and a novel regulator of mitosis in flies.
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the zinc finger protein asciz regulates b cell development via DYNLL1 and bim
Journal of Experimental Medicine, 2012Co-Authors: Sabine Jurado, Andreas Strasser, David M. Tarlinton, Kimberly Gleeson, David J. Izon, Carl R. Walkley, Kristy Odonnell, Joerg HeierhorstAbstract:Developing B lymphocytes expressing defective or autoreactive pre-B or B cell receptors (BCRs) are eliminated by programmed cell death, but how the balance between death and survival signals is regulated to prevent immunodeficiency and autoimmunity remains incompletely understood. In this study, we show that absence of the essential ATM (ataxia telangiectasia mutated) substrate Chk2-interacting Zn2+-finger protein (ASCIZ; also known as ATMIN/ZNF822), a protein with dual functions in the DNA damage response and as a transcription factor, leads to progressive cell loss from the pre-B stage onwards and severely diminished splenic B cell numbers in mice. This lymphopenia cannot be suppressed by deletion of p53 or complementation with a prearranged BCR, indicating that it is not caused by impaired DNA damage responses or defective V(D)J recombination. Instead, ASCIZ-deficient B cell precursors contain highly reduced levels of DYNLL1 (dynein light chain 1; LC8), a recently identified transcriptional target of ASCIZ, and normal B cell development can be restored by ectopic DYNLL1 expression. Remarkably, the B cell lymphopenia in the absence of ASCIZ can also be fully suppressed by deletion of the proapoptotic DYNLL1 target Bim. Our findings demonstrate a key role for ASCIZ in regulating the survival of developing B cells by activating DYNLL1 expression, which may then modulate Bim-dependent apoptosis.
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ATM Substrate Chk2-interacting Zn2+ Finger (ASCIZ) Is a Bi-functional Transcriptional Activator and Feedback Sensor in the Regulation of Dynein Light Chain (DYNLL1) Expression,
The Journal of biological chemistry, 2011Co-Authors: Sabine Jurado, Nora Tenis, Lindus A Conlan, Emma Baker, Nicolas C. Hoch, Kimberly Gleeson, Monique Smeets, David J. Izon, Joerg HeierhorstAbstract:The highly conserved DYNLL1 (LC8) protein was originally discovered as a light chain of the dynein motor complex, but is increasingly emerging as a sequence-specific regulator of protein dimerization with hundreds of targets and wide-ranging cellular functions. Despite its important roles, DYNLL1's own regulation remains poorly understood. Here we identify ASCIZ (ATMIN/ZNF822), an essential Zn2+ finger protein with dual roles in the DNA base damage response and as a developmental transcription factor, as a conserved regulator of DYNLL1 gene expression. DYNLL1 levels are reduced by ∼10-fold in the absence of ASCIZ in human, mouse and chicken cells. ASCIZ binds directly to the DYNLL1 promoter and regulates its activity in a Zn2+ finger-dependent manner. DYNLL1 protein in turn interacts with ten binding sites in the ASCIZ transcription activation domain, and high DYNLL1 levels inhibit the transcriptional activity of ASCIZ. In addition, DYNLL1 was also required for DNA damage-induced ASCIZ focus formation. The dual ability of ASCIZ to activate DYNLL1 gene expression and to sense free DYNLL1 protein levels enables a simple dynamic feedback loop to adjust DYNLL1 levels to cellular needs. The ASCIZ-DYNLL1 feedback loop represents a novel mechanism for auto-regulation of gene expression, where the gene product directly inhibits the transcriptional activator while bound at its own promoter.