[Effect of SH2A gene in cell signal transduction and its subcellular locolization].
OBJECTIVE
To examine the effect of SH2A gene in cell signal transduction and its subcellular localization.


METHODS
RT-PCR method was used to amplify the coding sequence of SH2A gene. Eukaryotic recombined expression vector pcDNA 3.1-SH2A was constructed, and then Bel7402 cell and COS7 cell transfected by liposome. Multiple kinase assay was performed to examine the activity of protein kinase (PKC), mitogen activated protein kinase (MAPK), tyrosine protein kinase (TPK) in the transfected cells. Meantime, pEGFP-SH2A vector was also constructed and the cells transfected with it were examined by fluorescent microscopy.


RESULTS
Recombined expression vector pcDNA3.2-SH2A and pEGFP-SH2A contained the coding sequence of SH2A cDNA. In both cell lines expressing SH2A gene, the cytoplasm PKC activity decreased by 40% or so, but no apparent alteration was found in MAPK and TPK activity. SH2A gene was found localized in the cytoplasm of transfected cells under fluorescent microscope.


CONCLUSIONS
SH2A gene may act as an inhibiting factor in PKC signal transduction, and it is localized in cytoplasm.
Ding
Qian
Q
Department of Medical Genetics, China Medical University, Shenyang, Liaoning 110001 PR China.
Zhao
Yan-yan
YY
Sun
Zhi-jun
ZJ
Yu
Da-hai
DH
D000818:Animals; D019556:COS Cells; D003593:Cytoplasm / Q000737:chemistry; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C474507:SH2D4A protein, human; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases