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Nucleic Acids Research, 1989, Vol. 17, No. 16 6591-6601
© 1989

MOLECULAR BIOLOGY

Aberrant expression of platelet-derived growth factor A-chaln cDNAs due to cryptic spiking of RNA transcripts In COS-1 cells

Robert J. Wise+, Stuart H. Orkin and Tucker Collins1,*

Departments of Hematoiogy -Oncology, Children's Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute Boston, MA 02115, USA 1Pathology, Brigham and Women's Hospital, Harvard Medical School Boston, MA 02115, USA

*To whom correspondence should be addressed

Received May 4, 1989. Revised July 21, 1989. Accepted July 21, 1989.

Platelet-derived growth factor (PDGF) is a cationic dinner 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 gene 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.

+ Present address: Genetics Institute, 87 Cambridge Park Drive, Cambridge, MA 02140, USA


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