ライフサイエンスコーパス: CstF

1 CstF-64 may, therefore, be absent in spermatocytes becau

2 CstF-64, the RNA-binding component of the cleavage stimu

3 through the cleavage stimulation factor 3' (CstF 3')-processing complex.

4 The cleavage stimulation factor-50 (CstF-50) has a role in this response, providing a link b

5 ified regions of the CstF subunits, CstF-77, CstF-64, and CstF-50, required for interaction with each

6 BRCA1 protein levels, increased amounts of a CstF/BARD1/BRCA1 complex were detected.

7 of the CstF subunits, CstF-77, CstF-64, and CstF-50, required for interaction with each other.

8 nly decreases the interaction with BARD1 and CstF, but also decreases the UV-induced inhibition of 3'

9 dance, and decreased association of CPSF and CstF subunits with the INTS6 locus.

10 the transcription unit, along with CPSF and CstF, during the initial stages of transcription, suppor

11 tinct sequence elements by CFI(m), CPSF, and CstF suggests that vertebrate poly(A) site definition is

12 Thus, loading of ELL2 and CstF-64 on RNA polymerase II was linked, caused enhanced

13 AA hexamer upstream of the cleavage site and CstF to a downstream GU-rich element.

14 lyadenylation under both LPS-stimulating and CstF-64-overexpressing conditions.

15 The BARD1-CstF-50 interaction inhibits polyadenylation in vitro.

16 e region downstream of a poly(A) site, block CstF-64 association with RNA, and inhibit the cleavage r

17 e mRNA polyadenylation, suggesting that both CstF-64 and tauCstF-64 function to inhibit polyadenylati

18 ognized by a heterotrimeric protein complex (CstF) through its 64 kDa subunit (CstF-64); the strength

19 entity and formation of a complex containing CstF-64, but not for trans-splicing.

20 ion experiments indicate that the Cdc73-CPSF-CstF complex is necessary for 3' mRNA processing in vitr

21 inding domain of the 64-kDa subunit of CstF (CstF-64) (64K RBD) is sufficient to define a functional

22 The 64 kDa subunit of CstF (CstF-64) contains an RNA binding domain and is responsib

23 elongation and 3' processing factors (DSIF, CstF-64, and CPSF-100) is also restored.

24 his observation, we disrupted the endogenous CstF-64 gene in the B cell line DT40 and replaced it wit

25 cells, giving us the opportunity to examine CstF-64 function in an isolated developmental system.

26 The mRNA polyadenylation factor CstF interacts with the BRCA1-associated protein BARD1,

27 by the heterotrimeric polyadenylation factor CstF, although how, and indeed if, all variations of thi

28 We next show that the polyadenylation factor CstF, plays a direct role in the DNA damage response.

29 ELL2 and the polyadenylation factor CstF-64 tracked together with RNA polymerase II across t

30 hat interact with the polyadenylation factor CstF-64, we uncovered an interaction with the transcript

31 on of PAF1-C with the polyadenylation factor CstF.

32 Ser2 phosphorylation, and processing factor CstF recruitment at wild-type and mutant IgM transgenes

33 ESCs that lack the 3' end-processing factor CstF-64.

34 induced manner with the 3' processing factor CstF-64.

35 ndance of the PA cleavage stimulatory factor CstF-64, the potent splicing suppressor PTB, and the hyp

36 unit of an essential polyadenylation factor (CstF-64) is specifically repressed in mouse primary B ce

37 of the G/U-rich cleavage stimulation factor (CstF) binding sites and the degenerate cleavage and poly

38 end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the po

39 ctor (CPSF) and cleavage stimulation factor (CstF) complexes that are required for the maturation of

40 Cleavage stimulation factor (CstF) is a heterotrimer necessary for the first step, en

41 Cleavage stimulation factor (CstF) is a heterotrimeric protein complex essential for

42 Cleavage stimulation factor (CstF) is one of the multiple factors required for mRNA p

43 ree subunits of cleavage stimulation factor (CstF) that is essential for mRNA polyadenylation.

44 subunits of the cleavage stimulation factor (CstF), and symplekin.

45 omponent of the cleavage stimulation factor (CstF), interacts with pre-mRNAs at sequences downstream

46 protein of the cleavage stimulation factor (CstF-64) in mouse male germ cells and in brain, a somati

47 protein of the cleavage stimulation factor (CstF-64) is altered in male germ cells, we examined its

48 subunit of the cleavage stimulation factor (CstF-64) recognizes GU-rich elements within the 3'-untra

49 protein of the cleavage stimulation factor (CstF-64), in mouse meiotic and postmeiotic germ cells.

50 anced binding of cleavage-stimulaton factor (CstF).

51 polyadenylation/cleavage stimulatory factor (CstF-64) increases 5-fold during the G0 to S phase trans

52 Here a polyadenylation factor, CstF-50 (cleavage stimulation factor), is shown to inter

53 yeast), and an mRNA polyadenylation factor, CstF-64 (Rna15 in yeast), and provided evidence that thi

54 subunit of the cleavage stimulation factor, CstF.

55 g sites for the cleavage stimulatory factor, CstF.

56 m a complex with the polyadenylation factors CstF and CPSF.

57 A Drosophila homologue for CstF-64 has now been isolated, both through homology wit

58 howed that the Hinge domain is necessary for CstF-64 interaction with CstF-77 and consequent nuclear

59 Our data support a role for CstF dimerization in pre-mRNA 3' end processing.

60 n of the late high-affinity binding site for CstF into the early polyadenylation region significantly

61 d to contain a single high-affinity site for CstF, as well as one consensus hexanucleotide sequence.

62 Human CstF-77 is one of the three subunits of cleavage stimula

63 tition with the Rna15 (yeast analog of human CstF-64 protein) subunit of the processing complex.

64 The primary structure of the human CstF-64 polyadenylation factor contains 12 nearly identi

65 In CstF-64, a small region, highly conserved in metazoa, is

66 In CstF-77, a proline-rich domain is necessary not only for

67 Reduction of tauCstF-64 in CstF-64-deficient ESCs results in even greater levels of

68 Strikingly, a 10-fold decrease in CstF-64 concentration did not markedly affect cell growt

69 ntaining hnRNP H, H', and F but deficient in CstF-64 in memory B-cell extracts but not in plasma cell

70 Although no changes were detected in CstF, BARD1, and BRCA1 protein levels, increased amounts

71 r levels of CstF-64 result in an increase in CstF trimer.

72 us protein A) induces no further increase in CstF-64 above that seen for proliferation alone.

73 We observed a 5-fold increase in CstF-64 expression following LPS treatment of RAW macrop

74 at occurred concomitant with the increase in CstF-64 expression.

75 Therefore, the increase in CstF-64 is associated with the G0 to S phase transition.

76 Therefore, an increase in CstF-64 levels is not necessary to mediate the different

77 The increase in CstF-64 protein was specific in that several other facto

78 ing in G0, show a similar 5-fold increase in CstF-64 when cultured under conditions inducing prolifer

79 The rise in CstF-64 is therefore insufficient to induce secretion.

80 The rise in CstF-64 occurs at a time when the amount of poly(A)-cont

81 We show that multiple factors, including CstF, cleavage-polyadenylation specificity factor, and s

82 with surprisingly high affinities, by intact CstF and were functional in reconstituted, CstF-dependen

83 CstF-64 is limiting for formation of intact CstF, that CstF has a higher affinity for the microm pol

84 BARD1, like CstF-50, also interacts with RNA polymerase II.

85 ody formation causes suppression of X-linked CstF-64 expression during pachynema.

86 uCstF-64 only partially compensates for lost CstF-64 function, despite being recruited to the histone

87 native polyadenylation mechanism to modulate CstF-77, highlighting the importance of the regulation o

88 y experiments, we found that human and mouse CstF-77 genes also contain an intronic poly(A) site, whi

89 To determine why the 64,000 Mr CstF-64 is not expressed in spermatocytes, we mapped its

90 Expression of the approximately 70,000 Mr CstF-64 was limited to meiotic spermatocytes and postmei

91 made dependent upon co-expression of an MS2-CstF-64 fusion protein.

92 ure of the HAT (half a TPR) domain of murine CstF-77, as well as its C-terminal subdomain.

93 Strikingly, this leads to a novel CstF-dependant enhancement of the poly(A) synthesis phas

94 In memory B cells the activity of CstF-64 binding to pre-mRNA, but not its amount, was red

95 responsible for the RNA binding activity of CstF.

96 Alignment of CstF-64 homologues shows that the proteins have a conser

97 sing in vitro by stabilizing the assembly of CstF on the core downstream URE.

98 processing by stabilizing the association of CstF with the RNA substrate.

99 Because augmentation of CstF-64 levels is neither necessary nor sufficient for I

100 table structure which helps focus binding of CstF to the core downstream URE.

101 because the mutant SRp20 inhibits binding of CstF to the exon 4 poly(A) site.

102 Overexpression of the CTD-binding domain of CstF p50 had a dominant-negative effect on 3' processing

103 t the structure of the RNA-binding domain of CstF-64 containing an RNA recognition motif (RRM) augmen

104 ture of the RNA-binding N-terminal domain of CstF-64 showed how the N-terminal RNA recognition motif

105 e show by NMR that the C-terminal domains of CstF-64 and Rna15 fold into a three-helix bundle with an

106 (gene symbol Cstf2t), which is a homolog of CstF-64 fitting the criteria we expected for the variant

107 g the RBD from the apparent yeast homolog of CstF-64, RNA15.

108 ressor of forked gene encodes a homologue of CstF-77, and mutations in it affect mRNA 3' end formatio

109 Cells with reduced levels of CstF display decreased viability following UV treatment,

110 d to induce a 40% reduction in the levels of CstF subunits, which may contribute to the increased rea

111 macrophages revealed that elevated levels of CstF-64 altered the expression of 51 genes, 14 of which

112 that the physiologically increased levels of CstF-64 observed in LPS-stimulated RAW macrophages contr

113 Higher levels of CstF-64 result in an increase in CstF trimer.

114 how the N-terminal RNA recognition motif of CstF-64 recognizes GU-rich RNAs.

115 Cstf2t) is a testis-expressed orthologue of CstF-64.

116 e primary B cells and that overexpression of CstF-64 is sufficient to switch heavy chain expression f

117 ESCs also express the tauCstF-64 paralog of CstF-64.

118 tes deadenylation by PARN in the presence of CstF-50, and that CstF-50/BARD1 can revert the cap-bindi

119 is known about the RNA binding properties of CstF, the protein-protein interactions required for its

120 el of cross-linking of the 64 kDa protein of CstF to polyadenylation substrate RNAs.

121 This directs the recruitment of CstF (cleavage stimulatory factor) to the terminator and

122 lighting the importance of the regulation of CstF-77 in various species.

123 RNA binding domain of the 64-kDa subunit of CstF (CstF-64) (64K RBD) is sufficient to define a funct

124 The 64 kDa subunit of CstF (CstF-64) contains an RNA binding domain and is res

125 interactions with the other two subunits of CstF as well as with other components of the polyadenyla

126 say that allows structure-function assays on CstF-64, a protein that binds to pre-mRNAs downstream of

127 Microarray analysis performed on CstF-64 overexpressing RAW macrophages revealed that ele

128 The concentration of one CstF subunit (CstF-64) increases during activation of B

129 Yeast Rna15 and its vertebrate orthologue CstF-64 play critical roles in mRNA 3 '-end processing a

130 uggesting that nuclear import of a preformed CstF complex is an essential step in polyadenylation.

131 acts containing the 3' end formation protein CstF-64 and the SL2 snRNP.

132 ponsible for interactions with two proteins, CstF-77 and symplekin, a nuclear protein of previously u

133 t CstF and were functional in reconstituted, CstF-dependent cleavage assays.

134 We show here that SLAP accurately reflects CstF-64-dependent polyadenylation, confirming the validi

135 site, which can be utilized to produce short CstF-77 transcripts lacking sequences encoding domains t

136 nd U-rich sequences are variants of a single CstF recognition motif.

137 also related to the human and mouse somatic CstF-64 (74.9% and 73.4% identity, respectively).

138 We localized the gene for the somatic CstF-64 to the X chromosome, which would be inactivated

139 By extension, the testis-specific CstF-64 may be expressed from an autosomal homolog of th

140 tocytes, suggesting that the testis-specific CstF-64 might control expression of meiosis-specific gen

141 Consecutive Us are required for a strong CstF-GU interaction and we show how UU dinucleotides are

142 n in the AAUAAA sequence, an inserted strong CstF binding site, an inserted simian virus 40 (SV40) la

143 in lines containing genomes with the strong CstF site or the late SV40 signal mutations, while a sig

144 he recombinant genomes containing the strong CstF site or the late SV40 signal, suggesting that alter

145 The concentration of one CstF subunit (CstF-64) increases during activation of B cells, and thi

146 n complex (CstF) through its 64 kDa subunit (CstF-64); the strength of this interaction affects the e

147 The 77 kDa subunit, CstF-77, is known to mediate interactions with the other

148 rst identified regions of the CstF subunits, CstF-77, CstF-64, and CstF-50, required for interaction

149 We have named the variant form "tau CstF-64," and we describe here the cloning and character

150 hat the activity of betaCstF-64 is less than CstF-64 on a strong polyadenylation signal, suggesting p

151 by PARN in the presence of CstF-50, and that CstF-50/BARD1 can revert the cap-binding protein-80 (CBP

152 limiting for formation of intact CstF, that CstF has a higher affinity for the microm poly(A) site t

153 Using SLAP, we determined that CstF-64 domains involved in RNA binding, interaction wit

154 Our results indicate that CstF plays an active role in the response to DNA damage,

155 Our results indicate that CstF-64 plays a key role in regulating IgM heavy chain e

156 Our results indicate that CstF-64 plays unexpected roles in regulating gene expres

157 Furthermore, we show that CstF, RNAP II and BARD1 are all found at sites of repair

158 In this study, we show that CstF-50 interacts with nuclear poly(A)-specific ribonucl

159 We further show that CstF-64 is limiting for formation of intact CstF, that C

160 These results suggest that CstF-64 plays a key role in modulating the cell cycle in

161 m of the AAUAAA in pre-mRNA, suggesting that CstF-64 and the hnRNPs compete for a similar region.

162 The CstF-50/PARN complex formation has a role in the inhibit

163 ulates the interaction between TFIIB and the CstF-64 component of the CstF 3' cleavage and polyadenyl

164 n with the second half likely to contain the CstF-77 interaction domain; a central region variable in

165 fect on 3' processing without disrupting the CstF complex.

166 In Drosophila, the CstF-64 gene has a single 63 bp intron, is transcribed t

167 nt 12-base U-rich sequence distinct from the CstF-64 consensus was identified.

168 d protein occurs but without a change in the CstF-64 level.

169 tween TFIIB and the CstF-64 component of the CstF 3' cleavage and polyadenylation complex.

170 e terminator and also the recruitment of the CstF and CPSF (cleavage and polyadenylation specific fac

171 We therefore first identified regions of the CstF subunits, CstF-77, CstF-64, and CstF-50, required f

172 ed family of neuronal splice variants of the CstF-64 mRNA, betaCstF-64, that we hypothesized to funct

173 Furthermore, forced overexpression of the CstF-64 protein also induced alternative poly(A) site se

174 We investigated by NMR the dynamics of the CstF-64 RNA-binding domain, both free and bound to two G

175 ing domains that are involved in many of the CstF-77 functions.

176 in, as evidenced by its association with the CstF complex, and by its ability to stimulate polyadenyl

177 so provide evidence that PARN along with the CstF/BARD1 complex participates in the regulation of end

178 tion in BARD1 (Gln564His) reduced binding to CstF and abrogated inhibition of polyadenylation.

179 of the HAT-N and HAT-C domains are unique to CstF-77.

180 ing the criteria we expected for the variant CstF-64 protein.

181 This suggested that the variant CstF-64 was an autosomal homolog activated during that t

182 -64 protein fits the criteria of the variant CstF-64, including antibody reactivity, size, germ cell

183 found that p53 can coexist in complexes with CstF and BARD1 in extracts of UV-treated cells, suggesti

184 2 snRNP still permits complex formation with CstF-64.

185 est that PC4/Sub1p, via its interaction with CstF-64/Rna15p, possesses an evolutionarily conserved an

186 ns involved in RNA binding, interaction with CstF-77 (the "Hinge" domain), and coupling to transcript

187 in is necessary for CstF-64 interaction with CstF-77 and consequent nuclear localization, suggesting

188 demonstrate that p53 directly interacts with CstF independent of TFIIB phosphorylation, providing an

189 It interacts with CstF-77, which in turn interacts with CPSF.

190 nt changes in the region that interacts with CstF-77.

191 utations prevent association of SL2 RNA with CstF-64.