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Nuclear translocation of prolactin: Collaboration of tyrosine kinase and protein kinase C activation in rat Nb2 node lymphoma cells
Author(s) -
Rao YiPing,
Buckley Donna J.,
Olson Mark D.,
Buckley Arthur R.
Publication year - 1995
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041630207
Subject(s) - internalization , protein kinase c , biology , chromosomal translocation , microbiology and biotechnology , tyrosine kinase , signal transduction , receptor , prolactin , calphostin c , autocrine signalling , kinase , cell culture , endocrinology , biochemistry , hormone , gene , genetics
Abstract Recent evidence has suggested that prolactin (PRL), internalized by lactogen‐dependent Nb2 lymphoma cells, is actively translocated to the nucleus where it binds to PRL receptors. Moreover, the mitogenic action of PRL in these cells has been separately linked to protein tyrosyl phosphorylation and activation of protein kinase C (PKC). Therefore, the coupling of PRL internalization and nuclear translocation to the activation of these signal transduction pathways was investigated. Results from control experiments indicated that 30% of internalized and 5% total cell‐associated 125 I‐rat PRL could be recovered within nuclei obtained from Nb2 cells previously incubated with the radiolabel for 3 h at 37°C. Furthermore, internalized PRL was found to be intact and not associated with any carrier proteins. Addition of tyrosine kinase (TK) antagonists, genistein or tyrphostin, significantly reduced cell surface binding, internalization, and nuclear translocation of 125 I‐rat PRL. In contrast, neither the level of cell‐associated nor internalized hormone differed between cells treated with the PKC antagonists, staurosporine or calphostin C, and control cultures. Instead, PKC inhibition significantly reduced nuclear PRL translocation. The inhibitory effects of the TK and PKC antagonists on PRL internalization and nuclear translocation in intact Nb2 cells were verified by immunofluorescence microscopy in parallel experiments. In other experiments, each of the kinase inhibitors blocked PRL‐stimulated Nb2 cell proliferation in a concentration‐dependent manner. It is concluded that activated TK and PKC collaborate in the process of PRL internalization and translocation to the nucleus. TK activation may participate in PRL receptor binding or hormone internalization while activation of PKC appears to be required for its nuclear targeting. Since TK and PKC activation are required for lactogen‐stimulated Nb2 cell proliferation, we suggest that a component of the mitogenic pathway in these cells is a direct nuclear interaction of PRL. © 1995 Wiley‐Liss, Inc.