An invariant asparagine residue belonging to a highly conserved domain in all protein kinases is instrumental in the protein kinase activity of the v-mil gene product.
P100gag-mil, the translation product of the v-mil oncogene of MH2 is a protein kinase specific of serine/threonine residues. We report here that the P100gag-mil encoded by the MH2-Hd isolate displays a considerably reduced kinase activity in vitro. Construction of chimeric viruses and sequencing revealed that the lesion responsible for this reduced activity results from a single point mutation converting an asparagine residue at position 720 in fully active P100gag-mil kinase into serine in the P100gag-mil of MH2-Hd. Since this asparagine residue together with an invariant aspartate residue bracket a highly conserved 6 amino-acid region in all known protein kinases as well as in phosphotransferases of bacterial origin, our results indicate that integrity of this region is essential to enzymatic function and support the notion that it could be directly involved in ATP binding or phosphate transfer from ATP to kinase substrates.
Denhez
F
F
CNRS UA 04 1160/INSERM U 186, Institut Pasteur, Lille, France.
Galibert
F
F
Heimann
B
B
Dozier
C
C
Graf
T
T
Moelling
K
K
Stehelin
D
D
Ghysdael
J
J
D000818:Animals; D001216:Asparagine / Q000235:genetics*; D001354:Avian Leukosis Virus / Q000235:genetics*; D015153:Blotting, Western; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D015683:Gene Products, gag; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000378:metabolism; D009857:Oncogenes; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D012191:Retroviridae Proteins / Q000235:genetics / Q000378:metabolism
D015683:Gene Products, gag; D009856:Oncogene Proteins, Viral; D012191:Retroviridae Proteins; D001216:Asparagine; D011494:Protein Kinases