Depletion of mammalian CCR4b deadenylase triggers elevation of the p27Kip1 mRNA level and impairs cell growth.
The stability of mRNA influences the abundance of cellular transcripts and proteins. Deadenylases play critical roles in mRNA turnover and thus are important for the regulation of various biological events. Here, we report the identification and characterization of CCR4b/CNOT6L, which is homologous to yeast CCR4 mRNA deadenylase. CCR4b is localized mainly in the cytoplasm and displays deadenylase activity both in vitro and in vivo. CCR4b forms a multisubunit complex similar to the yeast CCR4-NOT complex. Suppression of CCR4b by RNA interference results in growth retardation of NIH 3T3 cells accompanied by elevation of both p27(Kip1) mRNA and p27(Kip1) protein. Reintroduction of wild-type CCR4b, but not mutant CCR4b lacking deadenylase activity, restores the growth of CCR4b-depleted NIH 3T3 cells. The data suggest that CCR4b regulates cell growth in a manner dependent on its deadenylase activity. We also show that p27(Kip1) mRNA is stabilized and its poly(A) tail is preserved in CCR4b-depleted cells. Our findings provide evidence that CCR4b deadenylase is a constituent of the mammalian CCR4-NOT complex and regulates the turnover rate of specific target mRNAs. Thus, CCR4b may be involved in various cellular events that include cell proliferation.
Morita
Masahiro
M
Division of Oncology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.
Suzuki
Toru
T
Nakamura
Takahisa
T
Yokoyama
Kazumasa
K
Miyasaka
Takashi
T
Yamamoto
Tadashi
T
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D049109:Cell Proliferation; D003001:Cloning, Molecular; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary; D005095:Exoribonucleases / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D011061:Poly A / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry / Q000172:deficiency*; D013347:Subcellular Fractions / Q000378:metabolism; D014018:Tissue Distribution
D018076:DNA, Complementary; D012333:RNA, Messenger; D050760:Cyclin-Dependent Kinase Inhibitor p27; D011061:Poly A; C521732:CNOT6L protein, human; C521731:Cnot6l protein, mouse; D005095:Exoribonucleases; D012260:Ribonucleases; C445084:mRNA deadenylase