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Alternative splicing of bovine growth hormone pre-mRNA in vitro

Sun, Qiang
http://rave.ohiolink.edu/etdc/view?acc_num=case1058216813

Abstract Details

1995, Doctor of Philosophy, Case Western Reserve University, Molecular Biology and Microbiology.
Bovine growth hormone (bGH) pre-mRNA contains five exons and four introns. The last intron (intron D) is alternatively retained in a small fraction of the cytosolic bGH mRNA in bovine pituitary. The splicing of bGH intron D is positively influenced by a 115 nucleotide FspI-PvuII (FP) fragment in the downstream exon 5. The FP sequence enhances bGH intron D splicing and functions as an exonic splicing enhancer (ESE). To study the molecular mechanism by which the bGH ESE activates bGH intron D splicing, an in vitro system has been developed. The bGH intron D is accurately spliced in vitro. Furthermore, splicing of bGH intron D in vitro also requires stimulation by the bGH ESE, faithfully reflecting the exon influence on splicing as observed in vivo. The bGH ESE promotes early splicing complex formation. The bGH ESE appears to affect splicing through interaction with trans-acting factor(s) since it competes with bGH intron D splicing in trans. UV cross-linking experiments identified a 35 kDa protein factor(s) binding specifically to the bGH ESE. Immunoprecipitation assays identified the general splicing factor SF2/ASF as being one of the 35 kDa splicing factors binding to the bGH ESE. SF2/ASF activates splicing of bGH intron D through specific interaction with the bGH ESE. hnRNP A1, which counteracts SF2/ASF in alternative 5′ splice site select ion, also counteracts SF2/ASF in bGH intron D splicing in vitro. The bGH ESE contains novel, purine-rich sequence elements which can functionally replace the bGH ESE and activate bGH intron D splicing. The bGH ESE is not required for splicing of a β-globin intron, suggesting it is only required for splicing of weak introns, but not strong introns. bGH intron D contains both suboptimal 5′ and suboptimal 3′ splice sites. The bGH ESE promotes bGH intron D splicing through 3′ splice site activation in vitro. The bGH exon 5 also appears to contain negative splicing element(s), suggesting that splicing of bGH intron D is under both positive and negative regulation.
Fritz Rottman (Advisor)
152 p.
Biology, Molecular
Alternative splicing bovine growth hormone pre-mRNA in vitro

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Citations

  • Sun, Q. (1995). Alternative splicing of bovine growth hormone pre-mRNA in vitro [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1058216813

    APA Style (7th edition)

  • Sun, Qiang. Alternative splicing of bovine growth hormone pre-mRNA in vitro. 1995. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1058216813.

    MLA Style (8th edition)

  • Sun, Qiang. "Alternative splicing of bovine growth hormone pre-mRNA in vitro." Doctoral dissertation, Case Western Reserve University, 1995. http://rave.ohiolink.edu/etdc/view?acc_num=case1058216813

    Chicago Manual of Style (17th edition)

case1058216813
457
© 1995, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.