The G11 gene located in the major histocompatibility complex encodes a novel nuclear serine/threonine protein kinase.
Protein kinases are involved in signal transduction pathways and play fundamental roles in the regulation of cell functions. Here we report that the gene G11 located in the human major histocompatibility complex encodes a novel Ser/Thr protein kinase. The G11 gene products of 41.5 and 30 kDa were expressed in insect cells using the baculovirus system and transiently in the mammalian cell line COS-7. It was found that after immunoprecipitation of the G11 polypeptides from recombinant baculovirus-infected insect cell lysates or transfected COS-7 cell lysates the immunoprecipitates contained a Mn2+-dependent protein kinase activity that phosphorylated alpha-casein at Ser/Thr residues and histone at Ser residues. Furthermore, mutation of the ATP-binding site by converting the invariant lysine in the catalytic domain (amino acid 317) to a proline resulted in the complete ablation of the enzyme activity. This was consistent with the observation that the G11 polypeptide can be covalently modified by the reactive ATP analogue 5'-p-fluorosulfonylbenzoyladenosine in the absence of ATP, and that this modification is prevented in the presence of 1 mM ATP, indicating that the kinase domain of the G11 polypeptide is capable of binding ATP. Immunofluorescence staining of transfected COS-7 cells transiently expressing G11 revealed that this novel Ser/Thr protein kinase is localized predominantly in the nucleus.
Gomez-Escobar
N
N
MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU United Kingdom.
Chou
C F
CF
Lin
W W
WW
Hsieh
S L
SL
Campbell
R D
RD
D000241:Adenosine / Q000031:analogs & derivatives; D000345:Affinity Labels; D000818:Animals; D019556:COS Cells; D002364:Caseins / Q000378:metabolism; D002451:Cell Compartmentation; D006657:Histones / Q000378:metabolism; D008285:Major Histocompatibility Complex; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins; D018411:Spodoptera
D000345:Affinity Labels; D002364:Caseins; D006657:Histones; D009687:Nuclear Proteins; D011994:Recombinant Proteins; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D017346:Protein Serine-Threonine Kinases; C087996:STK19 protein, human; D000241:Adenosine