Abstract:
:We previously showed that receptor-like protein tyrosine phosphatase (RPTP)-alpha inhibited insulin-increased prolactin gene transcription. Others suggested that RPTPalpha was a key intermediary between integrins and activation of Src. We present evidence that inhibition of insulin-increased prolactin gene transcription was secondary to RPTPalpha activation of Src, reflecting its role as mediator of integrin responses. Src kinase activity was increased in GH4 cells transiently or stably expressing RPTPalpha and cells plated on the integrin-alpha5beta1 ligand fibronectin. C-terminal Src kinase inactivated Src and blocked RPTPalpha inhibition of insulin-increased prolactin gene transcription. Expression of dominant-negative Src also prevented the RPTPalpha-mediated inhibition of insulin-increased prolactin gene expression. Low levels of a constitutively active Src mutant (SrcY/F) stimulated whereas higher expression levels of Src Y/F inhibited prolactin gene expression. Src-increased prolactin gene transcription was inhibited by expression of a blocking Rho-mutant (RhoN19), suggesting that Src acted through or required active Rho. Experiments with an activated Rho-mutant (RhoL63) demonstrated a biphasic activation/repression of prolactin gene transcription that was similar to the effect of Src. The effects of both Src and Rho were phosphatidylinositol 3-kinase dependent. Expression of SrcY/F or RhoL63 altered the actin cytoskeleton and morphology of GH4 cells. Taken together, these data suggest a physiological pathway from the cell matrix to increased prolactin gene transcription mediated by RPTPalpha/Src/Rho/phosphatidylinositol 3-kinase and cytoskeletal change that is additive with effects of insulin. Over activation of this pathway, however, caused extreme alteration of the cytoskeleton that blocked activation of the prolactin gene.
journal_name
Endocrinologyjournal_title
Endocrinologyauthors
Vulin AI,Jacob KK,Stanley FMdoi
10.1210/en.2004-1386subject
Has Abstractpub_date
2005-08-01 00:00:00pages
3535-46issue
8eissn
0013-7227issn
1945-7170pii
en.2004-1386journal_volume
146pub_type
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