Journal Article

Effect of a tyrosine 155 to phenylalanine mutation of protein kinase Cδ on the proliferative and tumorigenic properties of NIH 3T3 fibroblasts

Péter Ács, Maryam Beheshti, Zoltán Szállási, Luowei Li, Stuart H. Yuspa and Peter M. Blumberg

in Carcinogenesis

Volume 21, issue 5, pages 887-891
Published in print May 2000 | ISSN: 0143-3334
Published online May 2000 | e-ISSN: 1460-2180 | DOI: http://dx.doi.org/10.1093/carcin/21.5.887
Effect of a tyrosine 155 to phenylalanine mutation of protein kinase Cδ on the proliferative and tumorigenic properties of NIH 3T3 fibroblasts

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Tyrosine phosphorylation has emerged as an important mechanism in the regulation of enzyme function. In this paper, we describe a mutant of PKCδ altered at a single tyrosine residue which has the opposite effect compared with wild-type PKCδ on the growth characteristics of NIH 3T3 cells. Overexpression of wild-type PKCδ results in a decreased growth rate and a lower cell density at confluency. On the other hand, overexpression of PKCδ with a mutation from tyrosine to phenylalanine at position 155 results in a significantly higher rate of growth and a higher density at confluency compared with vector controls. Moreover, these cells are able to grow in soft agar and to form tumors in nude mice. In contrast to kinase negative PKC constructs, this mutant maintains in vitro kinase activity and shows a subcellular localization and a translocation pattern that are similar to those of the wild-type PKCδ. Whether the altered biological effect is due to the missing phosphorylation on tyrosine or the mutation from tyrosine to phenylalanine per se remains under investigation.

Keywords: AcMBP, acetylated myelin basic protein; DMEM, Dulbecco's modified Eagle's medium; ECL, enhanced chemiluminescence; PAGE, polyacrylamide gel electrophoresis; PDBu, phorbol 12, 13-dibutyrate; PDGF, platelet derived growth factor; PKC, protein kinase C; PMA, phorbol 12-myristate 13-acetate; PS, phosphatidylserine.

Journal Article.  4580 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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