TY - JOUR
T1 - Generation of a Temperature-Sensitive cSRC
AU - Koegl, Manfred
AU - Goldberg, Yves
AU - Courtneidge, Sara A.
PY - 1993
Y1 - 1993
N2 - We have created retroviruses encoding a temperature-sensitive variant of the cSrc protein, and of its activated allele cSrc(Y527F). Cells expressing the activated allele harboring this mutation displayed a transformed phenotype when grown at 34°, but not at 39°, as quantitated by measuring the susceptibility of the cells to contact inhibition and their ability to grow anchorage-independently at the two temperatures. The level of the mutant protein in cells grown at the high temperature was slightly reduced, and the in vitro kinase activity of the protein was hypersensitive to inactivation by heat treatment: the combination of these two defects probably explains the temperature-sensitive nature of the transformed phenotype. Surprisingly, the pattern of tyrosine phosphorylated proteins differed very little between the two temperatures, suggesting that critical substrates are few and of low abundance. The introduction of the same mutation into the wild-type allele of cSrc, Y527, also rendered the protein thermolabile. These ecotropic retroviruses should prove useful in assessing the function of cSrc in mammalian cells.
AB - We have created retroviruses encoding a temperature-sensitive variant of the cSrc protein, and of its activated allele cSrc(Y527F). Cells expressing the activated allele harboring this mutation displayed a transformed phenotype when grown at 34°, but not at 39°, as quantitated by measuring the susceptibility of the cells to contact inhibition and their ability to grow anchorage-independently at the two temperatures. The level of the mutant protein in cells grown at the high temperature was slightly reduced, and the in vitro kinase activity of the protein was hypersensitive to inactivation by heat treatment: the combination of these two defects probably explains the temperature-sensitive nature of the transformed phenotype. Surprisingly, the pattern of tyrosine phosphorylated proteins differed very little between the two temperatures, suggesting that critical substrates are few and of low abundance. The introduction of the same mutation into the wild-type allele of cSrc, Y527, also rendered the protein thermolabile. These ecotropic retroviruses should prove useful in assessing the function of cSrc in mammalian cells.
UR - http://www.scopus.com/inward/record.url?scp=0027236580&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027236580&partnerID=8YFLogxK
U2 - 10.1006/viro.1993.1491
DO - 10.1006/viro.1993.1491
M3 - Article
C2 - 7689281
AN - SCOPUS:0027236580
SN - 0042-6822
VL - 196
SP - 368
EP - 371
JO - Virology
JF - Virology
IS - 1
ER -