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Research Article

Csk suppression of Src involves movement of Csk to sites of Src activity.

B W Howell, J A Cooper
B W Howell
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
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J A Cooper
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
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DOI: 10.1128/MCB.14.8.5402
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ABSTRACT

Csk phosphorylates Src family members at a key regulatory tyrosine in the C-terminal tail and suppresses their activities. It is not known whether Csk activity is regulated. To examine the features of Csk required for Src suppression, we expressed Csk mutants in a cell line with a disrupted csk gene. Expression of wild-type Csk suppressed Src, but Csk with mutations in the SH2, SH3, and catalytic domains did not suppress Src. An SH3 deletion mutant of Csk was fully active against in vitro substrates, but two SH2 domain mutants were essentially inactive. Whereas Src repressed by Csk was predominantly perinuclear, the activated Src in cells lacking Csk was localized to structures resembling podosomes. Activated mutant Src was also in podosomes, even in the presence of Csk. When Src was not active, Csk was diffusely located in the cytosol, but when Src was active, Csk colocalized with activated Src to podosomes. Csk also localizes to podosomes of cells transformed by an activated Src that lacks the major tyrosine autophosphorylation site, suggesting that the relocalization of Csk is not a consequence of the binding of the Csk SH2 domain to phosphorylated Src. A catalytically inactive Csk mutant also localized with Src to podosomes, but SH3 and SH2 domain mutants did not, suggesting that the SH3 and SH2 domains are both necessary to target Csk to places where Src is active. The failure of the catalytically active SH3 mutant of Csk to regulate Src may be due to its inability to colocalize with active Src.

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Csk suppression of Src involves movement of Csk to sites of Src activity.
B W Howell, J A Cooper
Molecular and Cellular Biology Aug 1994, 14 (8) 5402-5411; DOI: 10.1128/MCB.14.8.5402

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Csk suppression of Src involves movement of Csk to sites of Src activity.
B W Howell, J A Cooper
Molecular and Cellular Biology Aug 1994, 14 (8) 5402-5411; DOI: 10.1128/MCB.14.8.5402
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