Increased phosphorylation of histone H1 in mouse fibroblasts transformed with oncogenes or constitutively active mitogen-activated protein kinase kinase.
We compared the nucleosomal organization, histone H1 subtypes, and histone H1 phosphorylated isoforms of ras-transformed and parental 10T1/2 mouse fibroblasts. In agreement with previous studies, we found that ras-transformed mouse fibroblasts have a less condensed chromatin structure than normal fibroblasts. ras-transformed and parental 10T1/2 cells had similar amounts of H1 subtypes, proteins that have a key role in the compaction of chromatin. However, labeling studies with 32P and Western blot experiments with an antiphosphorylated H1 antibody show that interphase ras-transformed cells have higher levels of phosphorylated H1 isoforms than parental cells. G1/S phase-arrested ras-transformed cells had higher amounts of phosphorylated H1 than G1/S phase-arrested parental cells. Mouse fibroblasts transformed with fes, mos, raf, myc, or constitutively active mitogen-activated protein (MAP) kinase kinase had increased levels of phosphorylated H1. These observations suggest that increased phosphorylation of H1 is one of the consequences of the persistent activation of the mitogen-activated protein kinase signal transduction pathway. Indirect immunofluorescent studies show that phosphorylated H1b is localized in centers of RNA splicing in the nucleus, suggesting that this modified H1 subtype is complexed to transcriptionally active chromatin.
Chadee
D N
DN
Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Canada.
Taylor
W R
WR
Hurta
R A
RA
Allis
C D
CD
Wright
J A
JA
Davie
J R
JR
D016475:3T3 Cells; D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D016259:Genes, myc; D011905:Genes, ras; D006657:Histones / Q000378:metabolism*; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases; D009857:Oncogenes; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics*; D014170:Transformation, Genetic
D002843:Chromatin; D006657:Histones; D011494:Protein Kinases; D020929:Mitogen-Activated Protein Kinase Kinases