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PMID: 2780289 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Aberrant expression of platelet-derived growth factor A-chain cDNAs due to cryptic splicing of RNA transcripts in COS-1 cells.

Nucleic acids research ·Vol. 17 ·No. 16 ·1989-08-25 ·Pages 6591-601

Wise RJ, Orkin SH, Collins T

Abstract

Platelet-derived growth factor (PDGF) is a cationic dimer composed of two chains, designated A and B. All three dimeric isotypes of PDGF, PDGF-AA, -AB and -BB, are biologically active but may have distinct functional activities. Two A-chain precursors which differ by the presence of a highly basic 15 amino acid C-terminal extension are derived from the A-chain by alternative RNA splicing. To compare the functional properties of these two different forms of the A-chain, expression vectors were generated in which the cDNAs were placed under the transcriptional control of a viral promoter (pSV2). Surprisingly, cryptic RNA splice donor sites were identified in both forms of the PDGF A-chain which modify the A-chain open reading frame and alter the structure of the expressed protein. Recognition of this phenomenon appears to explain the discrepancies between previous results regarding the secretory properties of the PDGF A-chain and may explain difficulties in expression vectors containing splice acceptor sites between the inserted sequence and the polyadenylation site.

MeSH Terms
Animals Base Sequence Cell Line Codon/genetics DNA/genetics,isolation & purification Genetic Vectors Humans Macromolecular Substances Molecular Sequence Data Oligonucleotide Probes Platelet-Derived Growth Factor/genetics Protein Biosynthesis RNA Splicing RNA, Messenger/genetics Restriction Mapping Transcription, Genetic Transfection Umbilical Veins/metabolism
Chemicals
Codon Macromolecular Substances Oligonucleotide Probes Platelet-Derived Growth Factor RNA, Messenger DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wise R J
Department of Hematology, Children's Hospital, Boston, MA.
Orkin S H
Collins T
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33 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.16.6591
Published
1989-08-25
Pages
6591-601
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318352
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.16.6591 PMC Full Text View journal details
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