Interaction of p58(PITSLRE), a G2/M-specific protein kinase, with cyclin D3.
The p58(PITSLRE) is a p34(cdc2)-related protein kinase that plays an important role in normal cell cycle progression. Elevated expression of p58(PITSLRE) in eukaryotic cells prevents them from undergoing normal cytokinesis and appears to delay them in late telophase. To investigate the molecular mechanism of p58(PITSLRE) action, we used the yeast two-hybrid system, screened a human fetal liver cDNA library, and identified cyclin D3 as an interacting partner of p58(PITSLRE). In vitro binding assay, in vivo coimmunoprecipitation, and immunofluorescence cell staining further confirmed the association of p58(PITSLRE) with cyclin D3. This binding was observed only in the G(2)/M phase but not in the G(1)/S phase of the cell cycle; meanwhile, no interaction between p110(PITSLRE) and cyclin D3 was observed in all the cell cycle. The overexpression of cyclin D3 in 7721 cells leads to an exclusively accumulation of p58(PITSLRE) in the nuclear region, affecting its cellular distribution. Histone H1 kinase activity of p58(PITSLRE) was greatly enhanced upon interaction with cyclin D3. Furthermore, kinase activity of p58(PITSLRE) was found to increase greatly in the presence of cyclin D3 using a specific substrate, beta-1,4-galactosyltransferase 1. These data provide a new clue to our understanding of the cellular function of p58(PITSLRE) and cyclin D3.
Zhang
Songwen
S
Gene Research Center, Fudan University Medical Center (Former Shanghai Medical University), Shanghai, People's Republic of China 200032.
Cai
Mingmei
M
Zhang
Si
S
Xu
Songli
S
Chen
She
S
Chen
Xiaoning
X
Chen
Chun
C
Gu
Jianxin
J
D056743:Cyclin D3; D018844:Cyclin-Dependent Kinases; D016213:Cyclins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D016195:G2 Phase; D006801:Humans; D008938:Mitosis; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques
C534696:CCND3 protein, human; D056743:Cyclin D3; D016213:Cyclins; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; C447612:CDK11a protein, human; D018844:Cyclin-Dependent Kinases