期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 291, 期 26, 页码 13410-13420出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M116.727735
关键词
chromatin; histone modification; phosphorylation; transcription; ubiquitylation (ubiquitination); FACT; PAF-C; SILAC; casein kinase II; quantitative proteomics
资金
- National Institutes of Health [R01 GM099714, R01 GM110058]
- Indiana University School of Medicine
- DePauw University
- University of North Carolina Lineberger Comprehensive Cancer Center Basic Mechanisms of Viral and Chemical Carcinogenesis Training Grant [5 T32 CA009156]
- summer undergraduate program for prospective physician scientists at the Indiana University School of Medicine
Using affinity purification MS approaches, we have identified a novel role for casein kinase II (CKII) in the modification of the polymerase associated factor complex (PAF-C). Our data indicate that the facilitates chromatin transcription complex (FACT) interacts with CKII and may facilitate PAF complex phosphorylation. Posttranslational modification analysis of affinity-isolated PAF-C shows extensive CKII phosphorylation of all five subunits of PAF-C, although CKII subunits were not detected as interacting partners. Consistent with this, recombinant CKII or FACT-associated CKII isolated from cells can phosphorylate PAF-C in vitro, whereas no intrinsic kinase activity was detected in PAF-C samples. Significantly, PAF-C purifications combined with stable isotope labeling in cells (SILAC) quantitation for PAF-C phosphorylation from wild-type and CKII temperature-sensitive strains (cka1 cka2-8) showed that PAF-C phosphorylation at consensus CKII sites is significantly reduced in cka1 cka2-8 strains. Consistent with a role of CKII in FACT and PAF-C function, we show that decreased CKII function in vivo results in decreased levels of histone H2B lysine 123 monoubiquitylation, a modification dependent on FACT and PAF-C. Taken together, our results define a coordinated role of CKII and FACT in the regulation of RNA polymerase II transcription through chromatin via phosphorylation of PAF-C.
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