• Login as Editor
    View Item 
    •   Yeshiva Academic Institutional Repository
    • Albert Einstein College of Medicine (AECOM)
    • Albert Einstein College of Medicine: Doctoral Dissertations
    • View Item
    •   Yeshiva Academic Institutional Repository
    • Albert Einstein College of Medicine (AECOM)
    • Albert Einstein College of Medicine: Doctoral Dissertations
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Cloning and expression of a bovine cDNA for the regulatory subunit (RIIbeta) of cAMP-dependent protein kinase and cloning and characterization of the mouse RIIbeta gene

    Thumbnail

    Date
    1991
    Author
    Luo, Zhijun
    Metadata
    Show full item record
    Share
    Abstract
    cDNAs for RII{dollar}\beta{dollar} were cloned from a bovine brain {dollar}\lambda{dollar}gt11 library. The composite cDNA coded for a protein of 418 amino acids which was 98% homologous to the rat and human RII{dollar}\beta{dollar} proteins. The binding sites in RII{dollar}\beta{dollar} for microtubule-associated protein 2 (MAP2) and 75,000-dalton calmodulin-binding protein (P75) were localized to the NH{dollar}\sb2{dollar}-terminal 50 amino acids by using truncated RII{dollar}\beta{dollar} fusion proteins expressed in Escherichia Coli. MAP2 prevented RII from binding to P75 suggesting that the binding sites for MAP2 and P75 are located near each other or that one site on RII is binding to both proteins.;Genomic DNA clones for RII{dollar}\beta{dollar} were isolated from a mouse library in {dollar}\lambda{dollar}EMBL-3. 1.4 kb of the 5{dollar}\sp\prime{dollar}-flanking region upstream of the ATG initiation codon, exon 1 and part of intron 1 were sequenced. The 5{dollar}\sp\prime{dollar}-flanking region lacked TATA and CAAT sites but contained GC rich boxes found in constitutively expressed housekeeping genes. It contained five Sp1 sites, one each, AP1 and AP2 consensus sequences and ten copies of binding motifs for the helix-turn-helix class of transcription factors. Expression of fusion genes containing the CAT structural gene and RII{dollar}\beta{dollar} DNA (bp-1426/-49) upstream of the ATG initiation codon showed a 20 fold higher level of expression in NB2a cells than in CHO cells. Basal promoter was localized between {dollar}-{dollar}291 to {dollar}-{dollar}121 relative to ATG start codon. Deletion analysis suggested the presence of an enhancer element and inhibitory sequences. Gel shift assays revealed that several regions of the 5{dollar}\sp\prime{dollar}-flanking DNA interacted with nuclear binding factors in sequence and tissue specific manners. The defined basal promoter bound at least two factors present in both NB2a and CHO cells, one that bound Sp1 sites, one that bound to HLH motif. Our results suggest that tissue specific factors present in certain cell types are responsible for the preferential expression of RII{dollar}\beta{dollar}.
    Permanent Link(s)
    https://ezproxy.yu.edu/login?url=http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqm&rft_dat=xri:pqdiss:9205380
    https://hdl.handle.net/20.500.12202/3397
    Citation
    Source: Dissertation Abstracts International, Volume: 52-09, Section: B, page: 4598.;Advisors: Jack Erlichman.
    *This is constructed from limited available data and may be imprecise.
    Collections
    • Albert Einstein College of Medicine: Doctoral Dissertations [1673]

    Yeshiva University Libraries copyright © 2021  DuraSpace
    YAIR Self-Deposit | YAIR User's Guide | Take Down Policy | Contact Us
    Yeshiva University
     

     

    Browse

    AllCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    Login as Editor

    Statistics

    View Usage Statistics

    Yeshiva University Libraries copyright © 2021  DuraSpace
    YAIR Self-Deposit | YAIR User's Guide | Take Down Policy | Contact Us
    Yeshiva University