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Dean R Tolan – One of the best experts on this subject based on the ideXlab platform.

  • struCture of human brain fruCtose 1 6 bis phosphate Aldolase linking isozyme struCture with funCtion
    Protein Science, 2009
    Co-Authors: Tracy L Arakaki, Dean R Tolan, John A Pezza, Michelle A Cronin, Christopher E Hopkins, Danna B Zimmer, Karen N Allen
    Abstract:

    FruCtose-1,6-(bis)phosphate Aldolase is a ubiquitous enzyme that Catalyzes the reversible aldol Cleavage of fruCtose-1,6-(bis)phosphate and fruCtose 1-phosphate to dihydroxyaCetone phosphate and either glyCeral-dehyde-3-phosphate or glyCeraldehyde, respeCtively. Vertebrate Aldolases exist as three isozymes with different tissue distributions and kinetiCs: Aldolase A (musCle and red blood Cell), Aldolase B (liver, kidney, and small intestine), and Aldolase C (brain and neuronal tissue). The struCtures of human Aldolases A and B are known and herein we report the first struCture of the human Aldolase C, solved by X-ray Crystallography at 3.0 A resolution. StruCtural differenCes between the isozymes were expeCted to aCCount for isozyme-speCifiC aCtivity. However, the struCtures of isozymes A, B, and C are the same in their overall fold and aCtive site struCture. The subtle Changes observed in aCtive site residues Arg42, Lys146, and Arg303 are insuffiCient to Completely aCCount for the tissue-speCifiC isozymiC differenCes. Consequently, the struCtural analysis has been extended to the isozyme-speCifiC residues (ISRs), those residues Conserved among paralogs. A Complete analysis of the ISRs in the Context of this struCture demonstrates that in several Cases an amino aCid residue that is Conserved among Aldolase C orthologs prevents an interaCtion that oCCurs in paralogs. In addition, the struCture Confirms the Clustering of ISRs into disCrete patChes on the surfaCe and reveals the existenCe in Aldolase C of a patCh of eleCtronegative residues loCalized near the C terminus. Together, these struCtural Changes highlight the differenCes required for the tissue and kinetiC speCifiCity among Aldolase isozymes.

  • IdentifiCation of Neuronal Isozyme SpeCifiC Residues by Comparison of Goldfish Aldolase C to Other Aldolases
    Comparative Biochemistry and Physiology Part A: Physiology, 1997
    Co-Authors: Tanya Z Berardini, Mariola Drygas-williams, Gloria V. Callard, Dean R Tolan
    Abstract:

    AbstraCt A 2061 bp CDNA enCoding a goldfish (Carassius auratus) Aldolase was isolated from a goldfish brain library. The deduCed 362 amino aCid sequenCe is more similar to vertebrate brain (Aldolase C) and musCle Aldolases (Aldolase A) than to the liver isozymes (Aldolase B). Northern blot analysis indiCates strong expression of the mRNA in brain but not in liver or musCle, whiCh indiCates that this is Aldolase C rather than Aldolase A. Analysis of all known vertebrate Aldolase amino aCid sequenCes reveals five residues; Leu-57, Arg-314, Thr-324, Glu-332, and Gly-350 that are present exClusively in Aldolase Cs. The goldfish Clone possesses all five residues. The residues are primarily loCated in the Carboxyl-terminal region of the enzyme and may play a role in determining the neuronal isozyme-speCifiC properties of the enzyme. Furthermore, the existenCe of an Aldolase C in a teleost fish has impliCations with respeCt to the timing of genome dupliCation events that are thought to have been CritiCal in vertebrate evolution.

  • NonCoordinate Changes in the steady‐state mrna expressed from Aldolase A and Aldolase C genes during differentiation of ChiCken myoblasts
    Journal of Cellular Biochemistry, 1995
    Co-Authors: Rachel L. Meighan-mantha, Dean R Tolan
    Abstract:

    : In ChiCkens, as in all vertebrates, tissue-speCifiC expression of Aldolase isozymes A, B, and C is developmentally Coordinated. These developmental transitions in Aldolase expression have been studied most extensively by Charting enzyme aCtivity during normal and abnormal development of speCifiC vertebrate tissues. Indeed, Aldolase expression has been a key marker for normal differentiation and for retrodifferentiation during CarCinogenesis. Aldolase expression during ChiCken myoblast differentiation offers a model for investigating the regulatory meChanisms of these developmental transitions at the level of gene expression. For these studies, CDNAs enCoding the most isozyme-speCifiC regions of both ChiCken Aldolase A and C were Cloned. The ChiCken Aldolase A CDNA represents the first report of this sequenCe. Aldolase steady-state mRNA expression was measured during ChiCken myoblast differentiation in primary Cultures using RNase proteCtion assays with CRNA probes generated from these Aldolase CDNA Clones. Steady-state mRNA for Aldolase C, the predominant embryoniC Aldolase isozyme in ChiCkens, did not signifiCantly Change throughout myoblast differentiation. In Contrast, expression of steady-state mRNA for Aldolase A, the only Aldolase isozyme found in adult-skeletal musCle, was not deteCted until after myoblast fusion was approximately 50% Completed. Aldolase A expression gradually inCreased throughout myoblast differentiation until approximately 48 h after fusion was Completed when there was a dramatiC inCrease. These results are Contrasted with those of Turner et al. (1974) [Dev Biol 37:63-89] that showed a Coordinated switCh in isozyme aCtivities between the embryoniC Aldolase C and the musCle-speCifiC Aldolase A. This disCordant expression indiCates that the Aldolase A and C genes may employ different regulatory meChanisms during myoblast differentiation.

Karl Herrup – One of the best experts on this subject based on the ideXlab platform.

  • Purkinje Cell expression of the mouse Aldolase C gene in transgeniC miCe is direCted by an upstream regulatory element
    Brain research. Molecular brain research, 2005
    Co-Authors: Rita R. Romito-digiacomo, Ernst U. Walther, Elizabeth A. Williams, Karl Herrup
    Abstract:

    We have sought to understand the regulation of the expression pattern of Aldolase C (Zebrin II) in Cerebellar Purkinje Cells. Normally, Aldolase C is expressed in a series of sagittal stripes of Purkinje Cells interrupted by stripes of little or no expression. GenomiC Aldolase C:LaCZ fusion genes with 1.8 kb of sequenCe 5′ to the transCription start site drive CNS expression of LaCZ only in astroCytes and Cells of the pia mater. If the 5′ portion of the transgene is extended to a full 5.0 kb, expression is reliably observed in Purkinje Cells, yet none of the astroCyte expression is lost. We broke the additional 3.0 kb into 1.0 kb fragments and tested eaCh for Purkinje Cell enhanCer aCtivity when appended to the original 1.8 kb ConstruCt. We show that the 886 bp region from nuCleotide -2796 to -3682 (relative to the start of transCription) Contains virtually all of the Purkinje Cell enhanCer aCtivity. However, neither the full 5.0 kb nor the 886 bp region direCted a striped expression pattern, as is seen for the endogenous gene. Taken together, our study loCalizes a Purkinje Cell enhanCer to a small 5′ region of the Aldolase C gene and illustrates that the element(s) responsible for the normal anatomiCally Complex pattern of Aldolase C expression are separate from those Conferring Cell-type speCifiCity. The relationship of these findings to previous work in other laboratories is disCussed.

  • GenomiC sequenCes of Aldolase C (Zebrin II) direCt laCZ expression exClusively in non-neuronal Cells of transgeniC miCe
    Proceedings of the National Academy of Sciences of the United States of America, 1998
    Co-Authors: Ernst U. Walther, Richard Hawkes, Suzan Dziennis, Martin Dichgans, Stephen M. Maricich, Rita R. Romito, Fei Yang, Saul L. Zackson, Karl Herrup
    Abstract:

    Aldolase C is regarded as the brain-speCifiC form of fruCtose-1,6-bisphosphate Aldolase whereas Aldolase A is regarded as musCle-speCifiC. In situ hybridization of mouse Central nervous system using isozyme-speCifiC probes revealed that Aldolase A and C are expressed in Complementary Cell types. With the exCeption of Cerebellar Purkinje Cells, Aldolase A mRNA is found in neurons; Aldolase C message is deteCted in astroCytes, some Cells of the pia mater, and Purkinje Cells. We isolated Aldolase C genomiC Clones that span the entire protein Coding region from 1.5 kb 5′ to the transCription start site to 0.5 kb 3′ to the end of the last exon. The baCterial gene, laCZ, was inserted in two different loCations and the ConstruCts tested in transgeniC miCe. When the protein Coding sequenCes were replaCed with laCZ, three of five transgeniC lines expressed β-galaCtosidase only in Cells of the pia mater; one line also expressed in astroCyte-like Cells. When laCZ was inserted into the final exon (and all struCtural gene sequenCes were retained) transgene expression was observed in astroCytes in all regions of the Central nervous system as well as in pial Cells. Thus, with the exCeption of Purkinje Cell expression, the behavior of the full-length transgene mimiCs the endogenous Aldolase C gene. The results with the shorter transgene suggest that additional enhanCer elements exist within the intrageniC sequenCes. The absenCe of Purkinje Cell staining suggests that the Cis elements required for this expression must be loCated outside of the sequenCes used in this study.

  • Aldolase C/zebrin II and the regionalization of the Cerebellum
    Journal of Molecular Neuroscience, 1995
    Co-Authors: Richard Hawkes, Karl Herrup
    Abstract:

    The Cerebellum is Comprised of multiple bands of Cells, eaCh with CharaCteristiC afferent and efferent projeCtions, and patterns of gene expression. The most studied example of a striped pattern of expression is the antigen reCognized by monoClonal antibody antizebrin II. Zebrin II is expressed by subsets of Purkinje Cells that form an array of parasagittal bands that extend rostroCaudally throughout the Cerebellar Cortex, separated by similar bands of Purkinje Cells that do not express zebrin II. ReCent Cloning studies have revealed that the zebrin II antigen is the respiratory isoenzyme Aldolase C. This artiCle reviews the Cellular and moleCular Compartmentation of the Cerebellum together with the moleCular biology of the Aldolase C gene, and speCulates on possible reasons for a striped pattern of expression.

Richard Hawkes – One of the best experts on this subject based on the ideXlab platform.

  • zebrin ii Aldolase C expression in the Cerebellum of the western diamondbaCk rattlesnake Crotalus atrox
    PLOS ONE, 2015
    Co-Authors: Joel W Aspden, Carol L Armstrong, Cristian Gutierrezibanez, Richard Hawkes, Andrew N Iwaniuk, Tobias Kohl, David J Graham, Douglas R Wylie
    Abstract:

    Aldolase C, also known as Zebrin II (ZII), is a glyColytiC enzyme that is expressed in Cerebellar Purkinje Cells of the vertebrate Cerebellum. In both mammals and birds, ZII is expressed heterogeneously, suCh that there are sagittal stripes of Purkinje Cells with high ZII expression (ZII+), alternating with stripes of Purkinje Cells with little or no expression (ZII-). The patterns of ZII+ and ZII- stripes in the Cerebellum of birds and mammals are strikingly similar, suggesting that it may have first evolved in the stem reptiles. In this study, we examined the expression of ZII in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox). In Contrast to birds and mammals, the Cerebellum of the rattlesnake is muCh smaller and simpler, Consisting of a small, unfoliated dome of Cells. A pattern of alternating ZII+ and ZII- sagittal stripes Cells was not observed: rather all Purkinje Cells were ZII+. This suggests that ZII stripes have either been lost in snakes or that they evolved Convergently in birds and mammals.

  • Zebrin II / Aldolase C expression in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox).
    PLOS ONE, 2015
    Co-Authors: Joel W Aspden, Carol L Armstrong, Richard Hawkes, Andrew N Iwaniuk, Tobias Kohl, David J Graham, Cristián Gutiérrez-ibáñez, Douglas R Wylie
    Abstract:

    Aldolase C, also known as Zebrin II (ZII), is a glyColytiC enzyme that is expressed in Cerebellar Purkinje Cells of the vertebrate Cerebellum. In both mammals and birds, ZII is expressed heterogeneously, suCh that there are sagittal stripes of Purkinje Cells with high ZII expression (ZII+), alternating with stripes of Purkinje Cells with little or no expression (ZII-). The patterns of ZII+ and ZII- stripes in the Cerebellum of birds and mammals are strikingly similar, suggesting that it may have first evolved in the stem reptiles. In this study, we examined the expression of ZII in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox). In Contrast to birds and mammals, the Cerebellum of the rattlesnake is muCh smaller and simpler, Consisting of a small, unfoliated dome of Cells. A pattern of alternating ZII+ and ZII- sagittal stripes Cells was not observed: rather all Purkinje Cells were ZII+. This suggests that ZII stripes have either been lost in snakes or that they evolved Convergently in birds and mammals.

  • Aldolase C/zebrin II expression in the neonatal rat forebrain reveals Cellular heterogeneity within the subventriCular zone and early astroCyte differentiation.
    The Journal of neuroscience : the official journal of the Society for Neuroscience, 2001
    Co-Authors: Susan M. Staugaitis, Richard Hawkes, Marielba Zerlin, Joel M. Levine, James E. Goldman
    Abstract:

    During late gestational and early postnatal development, proliferating Cells in the subventriCular zones of the lateral ventriCles (SVZ) migrate into the gray and white matter of the forebrain and differentiate into astroCytes and oligodendroCytes. BeCause the Cellular Composition and struCture of the neonatal SVZ is poorly understood, we performed a differential display PCR sCreen to identify genes preferentially expressed therein. One highly expressed gene enCoded Aldolase C. We used a speCifiC monoClonal antibody, Aldolase C/zebrin II (ALDC/ZII), in Combination with markers of glial lineage and proliferation, to CharaCterize the Cells that express this gene. In the neonatal SVZ, ALDC/ZII-positive Cells, whiCh are generally polygonal and display several proCesses, have a nonuniform spatial distribution. They do not express vimentin, GFAP, or NG2. A subset of ALDC/ZII-positive Cells inCorporates bromodeoxyuridine, but progenitors identified by β-galaCtosidase expression after infeCtion with reCombinant BAG virus do not show ALDC/ZII immunoreaCtivity. Outside of the SVZ, β-galaCtosidase-positive/ALDC/ZII-positive Cells have an astroCytiC phenotype, suggesting that immunoreaCtivity was aCquired after exit from the SVZ. These studies demonstrate that the neonatal SVZ is Composed of different populations of Cells that Can be CharaCterized by their antigeniC phenotype, their proliferative CapaCity, and their spatial distributions. Nonrandom distributions of different Cell types within the SVZ may permit the formation of miCroenvironments that stimulate the produCtion of Cells with speCifiC potentials at appropriate points in development. Analysis of ALDC/ZII expression by astroCyte lineage Cells in the neonatal Cerebral Cortex and white matter may reveal insights into the phenotype and behavior of undifferentiated astroCyte progenitors.

Izumi Sugihara – One of the best experts on this subject based on the ideXlab platform.

  • The entire trajeCtories of single pontoCerebellar axons and their lobular and longitudinal terminal distribution patterns in multiple Aldolase C‐positive Compartments of the rat Cerebellar Cortex
    The Journal of comparative neurology, 2019
    Co-Authors: Izumi Sugihara, Yoshikazu Shinoda
    Abstract:

    The mammalian Cerebellar Cortex is Compartmentalized, both anatomiCally and histoChemiCally, into multiple parasagittal bands. To CharaCterize the multiple zonal patterns of pontoCerebellar mossy fiber projeCtion, single neurons in the basilar pontine nuCleus (BPN) were labeled by injeCting biotinylated dextran amine into the BPN, and the entire axonal trajeCtory of single labeled neurons (n = 25) was reConstruCted in relation to Aldolase C Compartments of Purkinje Cells in rats. Single pontoCerebellar axons, after passing through the Contralateral middle Cerebellar pedunCle, ran transversely in the deep Cerebellar white matter toward and often aCross the midline, and on their ways, gave rise to 2-10 primary Collaterals at almost right angles in speCifiC lobules only Contralaterally or bilaterally with Contralateral predominanCe. EaCh primary Collateral further branChed in a parasagittal plane to form a strip-shaped longitudinal termination zone with rosette-type swellings Clustered in Aldolase C-positive Compartments in a single or multiple lobules, mainly in Compartment 4+//5+, 5+//6+, and 6+//7+. Axons arising from the Central, rostral, and lateral part of the BPN projeCted with multiple branChes, mainly to simple lobule, Crus II and paramedian lobule, to Crus I and dorsal parafloCCulus, and to ventral parafloCCulus and lobule IXC, respeCtively. The results showed the pontoCerebellar projeCtion is Closely related to lobular and Compartmental organization of the Cerebellum. A Comparison of single axon morphologies of different mossy fiber systems indiCates that the projeCtion pattern of single pontoCerebellar neurons with multiple Collaterals innervating different longitudinal Compartments arranged in a mediolateral direCtion represents a general feature of mossy fiber projeCtion.

  • Single axonal morphology and termination to Cerebellar Aldolase C stripes CharaCterize distinCt spinoCerebellar projeCtion systems originating from the thoraCiC spinal Cord in the mouse.
    The Journal of comparative neurology, 2017
    Co-Authors: Yuanjun Luo, Radhika Pooja Patel, Gideon Anokye Sarpong, Kazuma Sasamura, Izumi Sugihara
    Abstract:

    The spinoCerebellar projeCtion has an essential role in sensorimotor Coordination of limbs and the trunk. Multiple groups of spinoCerebellar projeCtions have been identified in retrograde labeling studies. In this study, we aimed at CharaCterizing projeCtion patterns of these groups using a Combination of anterograde labeling of the thoraCiC spinal Cord and Aldolase C immunostaining of longitudinal stripes of the Cerebellar Cortex in the mouse. We reConstruCted 22 single spinoCerebellar axons, wholly in the Cerebellum and brain stem and partly, in the spinal Cord. They were Classified into three groups, (a) non-Crossed axons of Clarke’s Column neurons (NCC, 8 axons), (b) non-Crossed axons of marginal Clarke’s Column neurons (NMCC, 7 axons), and (C) Crossed axons of neurons in the medial ventral horn (CMVH, 7 axons), based on previous retrograde labeling studies. While NCC axons projeCted mainly to multiple bilateral stripes in vermal lobules II-IV and VIII-IX, and the ipsilateral medial Cerebellar nuCleus, NMCC axons projeCted mainly to ipsilateral stripes in paravermal lobules II-V and Copula pyramidis, and the anterior interposed nuCleus. CMVH axons projeCted bilaterally to multiple stripes in lobules II-V with a small number of terminals but had abundant Collaterals in the spinal Cord and medullary retiCular nuClei as well as in the vestibular and Cerebellar nuClei. The results indiCate that, while CMVH axons overlap with propriospinal and spinoretiCular projeCtions, NCC and NMCC axons are primarily spinoCerebellar axons, whiCh seem to be involved in relatively more proximal and distal sensorimotor Controls, respeCtively.

  • Detailed Expression Pattern of Aldolase C (AldoC) in the Cerebellum, Retina and Other Areas of the CNS Studied in AldoC-Venus KnoCk-In MiCe
    , 2016
    Co-Authors: Hirofumi Fujita, Maya Yamazaki, Kenji Sakimura, Arata Oh-nishi, Hanako Aoki, Itsuki Ajioka, Manabu Abe, Izumi Sugihara
    Abstract:

    Aldolase C (AldoC, also known as ‘‘zebrin II’’), a brain type isozyme of a glyColysis enzyme, is expressed heterogeneously in subpopulations of Cerebellar Purkinje Cells (PCs) that are arranged longitudinally in a Complex striped pattern in the Cerebellar Cortex, a pattern whiCh is Closely related to the topography of input and output axonal projeCtions. Here, we generated knoCk-in AldoC-Venus miCe in whiCh AldoC expression is visualized by expression of a fluoresCent protein, Venus. SinCe there was no obvious phenotypes in general brain morphology and in the striped pattern of the Cerebellum in mutants, we made detailed observation of AldoC expression pattern in the nervous system by using Venus expression in AldoC-Venus heterozygotes. High levels of Venus expression were observed in Cerebellar PCs, Cartwheel Cells in the dorsal CoChlear nuCleus, sensory epithelium of the inner ear and in all major types of retinal Cells, while moderate levels of Venus expression were observed in astroCytes and satellite Cells in the dorsal root ganglion. The striped arrangement of PCs that express Venus to different degrees was Carefully traCed with serial seCtion alignment analysis and mapped on the unfolded sCheme of the entire Cerebellar Cortex to re-identify all individual AldoC stripes. A longitudinally striped boundary of AldoC expression was first identified in the mouse floCCulus, and was Correlated with the Climbing fiber projeCtion pattern an

Henriette Skala – One of the best experts on this subject based on the ideXlab platform.

  • Upstream elements involved in vivo in aCtivation of the brain-speCifiC rat Aldolase C gene. Role of binding sites for POU and winged helix proteins.
    The Journal of biological chemistry, 1998
    Co-Authors: Henriette Skala, Muriel Thomas, Axel Kahn, Arlette Porteu, Marie-france Szajnert, Hitoshi Okazawa, F Phan-dinh-tuy
    Abstract:

    The rat Aldolase C gene enCodes a glyColytiC enzyme strongly expressed in adult brain. We previously reported that a 115-base pair (bp) promoter fragment was able to ensure the brain-speCifiC expression of the ChlorampheniCol aCetyltransferase (CAT) reporter gene in transgeniC miCe, but only at a low level (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H. (1993) Eur. J. BioChem. 218, 143-151). Here we show that in vivo aCtivation of this promoter at a high level requires Cooperation between an upstream 0.6-kilobase pair (kb) fragment and far upstream sequenCes. In the 0.6-kb region, a 28-bp DNA element is shown to inClude overlapping in vitro binding sites for POU domain regulatory proteins and for the Winged Helix hepatoCyte nuClear faCtor-3beta faCtor. An hepatoCyte nuClear faCtor-3beta-binding site previously desCribed in the short proximal promoter fragment is also shown to interaCt in vitro with POU proteins, although with a lower affinity than the 28-bp motif. Additional binding sites for POU faCtors were deteCted in the upstream 0.6-kb sequenCes. Progressive deletion in this region resulted in deCreased expression levels of the transgenes in miCe, suggesting synergistiC interaCtions between these multiple POU-binding sites. We propose that DNA elements CharaCterized by a dual binding speCifiCity for both POU domain and Winged Helix transCription faCtors Could play an essential role in the brain-speCifiC expression of the Aldolase C gene and other neuronal genes.

  • FunCtional DisseCtion of the Brain-speCifiC Rat Aldolase C Gene Promoter in TransgeniC MiCe: ESSENTIAL ROLE OF TWO GC-RICH BOXES AND AN HNF3 BINDING SITE (∗)
    The Journal of biological chemistry, 1995
    Co-Authors: Muriel Thomas, Henriette Skala, Axel Kahn, Françoise Phan Dinh Tuy
    Abstract:

    AbstraCt The Aldolase C gene produCt is a glyColytiC isoenzyme speCifiCally deteCted in brain. We have previously defined a short 115-base pair promoter fragment able to Confer on a reporter ChlorampheniCol aCetyltransferase (CAT) gene a speCifiC expression in brain of transgeniC miCe. In this promoter fragment, two GC-riCh regions (A/A‘ and B boxes) were deteCted by in vitro DNase1 footprinting experiments with brain, fibroblast, or liver nuClear extraCts. Both A/A‘ and B boxes, sharing struCtural homology, are able to interaCt with Sp1, Krox20/Krox24 faCtors and with other proteins (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H.(1993) Eur. J. BioChem. 218, 143-151). In this paper, we desCribe a new ubiquitous faCtor termed Ub able to bind the A/A‘ box. We also delimit a third element (box C) binding a hepatoCyte-enriChed protein displaCed by a hepatoCyte nuClear faCtor 3-speCifiC oligonuCleotide. The funCtional involvement of eaCh binding site in brain-speCifiC transCription of the Aldolase C gene has been tested in transgeniC miCe Carrying different mutant promoters Cloned in front of the CAT gene. A promoter Containing only box C was totally inaCtive, suggesting an essential role of the region Containing A/A‘ and B boxes. However, mutations or deletions of either the A/A‘ or the B box have no signifiCant effeCt on the CAT gene expression. We therefore hypothesize that the A/A‘ and B sites may be funCtionally redundant. Indeed, ConstruCts harboring only one of these two boxes (A/A‘ or B) linked to the C box displayed a brain-speCifiC CAT aCtivity similar to that obtained with the wild-type promoter. Furthermore, a transgene with disruption of the C box, keeping intaCt the A/A‘ and B boxes, was totally inaCtive, suggesting a CruCial role of the hepatoCyte nuClear faCtor 3 binding site in aCtivation of the Aldolase C gene.

  • FunCtional disseCtion of the brain-speCifiC rat Aldolase C gene promoter in transgeniC miCe
    Journal of Biological Chemistry, 1995
    Co-Authors: Muriel Thomas, Henriette Skala, Axel Kahn, Françoise Phan Dinh Tuy
    Abstract:

    The Aldolase C gene produCt is a glyColytiC isoenzyme speCifiCally deteCted in brain. We have previously defined a short 115-base pair promoter fragment able to Confer on a reporter ChlorampheniCol aCetyltransferase (CAT) gene a speCifiC expression in brain of transgeniC miCe. In this promoter fragment, two GC-riCh regions (A/A’ and B boxes) were deteCted by in vitro DNase1 footprinting experiments with brain, fibroblast, or liver nuClear extraCts. Both A/A’ and B boxes, sharing struCtural homology, are able to interaCt with Sp1, Krox20/Krox24 faCtors and with other proteins (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H. (1993) Eur. J. BioChem. 218, 143-151). In this paper, we desCribe a new ubiquitous faCtor termed Ub able to bind the A/A’ box. We also delimit a third element (box C) binding a hepatoCyte-enriChed protein displaCed by a hepatoCyte nuClear faCtor 3-speCifiC oligonuCleotide. The funCtional involvement of eaCh binding site in brain-speCifiC transCription of the Aldolase C gene has been tested in transgeniC miCe Carrying different mutant promoters Cloned in front of the CAT gene. A promoter Containing only box C was totally inaCtive, suggesting an essential role of the region Containing A/A’ and B boxes. However, mutations or deletions of either the A/A’ or the B box have no signifiCant effeCt on the CAT gene expression. We therefore hypothesize that the A/A’ and B sites may be funCtionally redundant. Indeed, ConstruCts harboring only one of these two boxes (A/A’ or B) linked to the C box displayed a brain-speCifiC CAT aCtivity similar to that obtained with the wild-type promoter. Furthermore, a transgene with disruption of the C box, keeping intaCt the A/A’ and B boxes, was totally inaCtive, suggesting a CruCial role of the hepatoCyte nuClear faCtor 3 binding site in aCtivation of the Aldolase C gene.