Functional asymmetry of the regions juxtaposed to the membrane-binding sequence of polyomavirus middle T antigen.

Abstract:

:The functional importance of the two clusters of positively charged amino acids which flank the hydrophobic membrane-anchoring sequence of polyomavirus middle T (mT) protein has been investigated by using site-directed mutagenesis. A clear asymmetry was apparent. No effect on transformation was seen following multiple alterations or complete removal of the cluster at the carboxyl end of the protein. In contrast, a single substitution replacing the first arginine amino terminal to the hydrophobic stretch with glutamic acid, but not with lysine, histidine, or methionine, produced a partially transformation-defective mutant with a novel phenotype. This mutant failed to confer anchorage-independent growth on F111 established rat embryo fibroblasts but induced foci with altered morphology compared with wild-type mT. Biochemical studies on this mutant revealed that F111 clones expressing levels of mutant mT equivalent to those of wild-type controls showed a 65% reduction in pp60c-src activation and an 87% reduction in mT-associated phosphatidylinositol 3-kinase activity. However, F111 clones expressing seven times more mutant mT than did wild-type controls showed equal or greater levels of kinase activities yet remained incompletely transformed. Possible mechanisms involving this transformation-sensitive region of mT are discussed.

journal_name

Mol Cell Biol

authors

Dahl J,Thathamangalam U,Freund R,Benjamin TL

doi

10.1128/mcb.12.11.5050

subject

Has Abstract

pub_date

1992-11-01 00:00:00

pages

5050-8

issue

11

eissn

0270-7306

issn

1098-5549

journal_volume

12

pub_type

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