CELL-CYCLE REENTRY OF MAMMALIAN FIBROBLASTS IS ACCOMPANIED BY THE SUSTAINED ACTIVATION OF P44(MAPK) AND P42(MAPK) ISOFORMS IN THE G1 PHASE AND THEIR INACTIVATION AT THE G1/S TRANSITION

Mitogen-activated protein (MAP) kinases are serine/threonine kinases that are rapidly activated in response to mitogenic stimuli. Here we examined the enzymatic activity and phosphorylation state of the individual p44(mapk) and p42(mapk) isoforms during early G1 and late G1 phase of the mammalian cell cycle. Release of fibroblast cells from early G1 block was accompanied by a rapid rise in the myelin basic protein (MBP) kinase activity of p44(mapk) and p42(mapk), which declined slowly over several hours to reach negligible values as cells enter S phase. When cells were released from late G1 block, the activity of p44(mapk) and p42(mapk) increased transiently, and then rapidly declined to baseline values during G1 to S phase transition. Cells released at the G1/S boundary in a medium lacking growth factors entered S phase in the complete absence of MAP kinase activity. Unlike MAP kinases, the histone H1 kinase activity of p33(cdk2) Was elevated in late G1-arrested cells and continued to increase during S phase entry. The enzymatic activation of p44(mapk) and p42(mapk) in both early G1 and late G1 phase was accompanied by an increase in the phosphothreonine and phosphotyrosine content of the proteins. These findings suggest that the sustained activation of MAP kinases during G1 progression and their inactivation at the G1/S transition are two regulatory processes involved in the mitogenic response to growth factors. (C) 1995 Wiley-Liss, Inc.