Sa. Ezhevsky et al., ROLE OF CYCLIN-A AND P27 IN ANTI-IGM-INDUCED G(1) GROWTH ARREST OF MURINE B-CELL LYMPHOMAS, Molecular biology of the cell, 7(4), 1996, pp. 553-564
Cross-linking surface immunoglobulin (Ig)M on the WEHI-231 B-cell lymp
homa results in decreased cell size, G(1)/S growth arrest, and finally
DNA cleavage into oligonucleosomal fragments that are the classical f
eatures of apoptotic cells. Treatment of WEHI-231 cells with anti-IgM
in early G(1) phase prevents phosphorylation of the retinoblastoma gen
e product (pRb) and inhibits entry into S phase. Using unsynchronized
cells, we previously demonstrated that cyclin A-associated and Cdk2-de
pendent GST-pRb kinase activity were inhibited in WEHI-231 cells treat
ed with anti-IgM. We now show that progression of elutriated early G(1
) phase WEHI-231 cells from early into late G(1) phase is accompanied
by an increase in the abundance of cyclin A protein and cyclin A-assoc
iated kinase activity. Treatment of early G(1) cells with anti-IgM pre
vented this increase in cyclin A-associated kinase activity at late G(
1), despite minimal changes in the overall level of cyclin A and Cdk2
proteins. Late G(1) cells, which already possess high cyclin A-associa
ted kinase activity, were insensitive to anti-IgM treatment and were a
ble to complete the cell cycle. We also found that anti-IgM-treated ce
lls contained increased amounts of the Cdk inhibitor protein p(27Kip1)
. Essentially all of the cyclin A in treated cells was associated with
p27, a result which we propose explains the lack of cyclin A/Cdk2 kin
ase activity. Accumulation of p27 in cyclin A kinase complexes, howeve
r, did not decrease the amount of Cdk2 bound to cyclin A. Thus, cross-
Linking IgM on growth-inhibitable B-cell lymphomas affects cyclin A ki
nase activity by increasing the levels of p27 in this complex, thus pr
eventing productive pRb phosphorylation and leading to cell cycle arre
st and subsequent apoptosis. These results are discussed in terms of t
he cell cycle restriction points that regulate lymphocyte function, as
well as the lineage-specific differences in cell cycle control.