Mapping by in vitro constructs of the P100gag-mil region, accounting for induction of chicken neuroretina cell proliferation.
The v-mil oncogene of the avian retrovirus MH2 is expressed as a fusion protein with viral gag determinants in infected cells. This P100gag-mil protein accounts for the proliferation of chicken embryo neuroretina cells (CNR) induced by MH2 in vitro. We constructed a series of mutants by in-frame deletions in different parts of the gag and mil domains and tested their ability to induce CNR growth. We show that gag sequences, as well as 200-base-pair 5' mil sequences, were not required to induce such a proliferation. However, gag sequences seem to contribute to a full proliferation of growing CNR. In contrast, deletions in the kinase domain abolish this induction. In particular, by deleting only 9 nucleotides localized around the unique SphI site of v-mil, we produced a totally inactive mutant (BalSp). This mutant directs the synthesis of a v-mil protein lacking the dipeptide Tyr-Leu, which is conserved in almost all the members of the large protein kinase family, and a histidine residue highly conserved in Ser-Thr protein kinase members.
Coll
J
J
Institut National de la Santé et de la Recherche Médicale U 186, Institut Pasteur de Lille, France.
Dozier
C
C
Saule
S
S
Henry
C
C
Quatannens
B
B
Debuire
B
B
Stehelin
D
D
D000818:Animals; D002453:Cell Cycle; D002478:Cells, Cultured; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D003370:Coturnix; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D015683:Gene Products, gag; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D009857:Oncogenes; D012160:Retina / Q000166:cytology* / Q000382:microbiology; D012191:Retroviridae Proteins / Q000235:genetics*; D013329:Structure-Activity Relationship; D014162:Transfection; D014779:Virus Replication
D015683:Gene Products, gag; D009856:Oncogene Proteins, Viral; D012191:Retroviridae Proteins