Aberrant expression of platelet-derived growth factor A-chain cDNAs due to cryptic splicing of RNA transcripts in COS-1 cells.
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.
Department of Hematology, Children's Hospital, Boston, MA.
R J
RJ
Wise
S H
SH
Orkin
T
T
Collins
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003062:Codon / Q000235:genetics; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D014176:Protein Biosynthesis; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection; D014471:Umbilical Veins / Q000378:metabolism
D003062:Codon; D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D004247:DNA