ライフサイエンスコーパス: U2AF(65)

1 ted from high affinity binding sequences for U2AF65.

2 interact with a U2AF homology motif (UHM) of U2AF65.

3 nal strong cross-link to the splicing factor U2AF65.

4 onical polypyrimidine tract binding protein, U2AF65.

5 ete, <20 nt poly(A) tail in cells expressing U2AF65.

6 laboratively with MUD2, the yeast homolog of U2AF65.

7 rp2 protein (spU2AF59), a homologue of human U2AF65.

8 nRNP as well as Mud2p, which resembles human U2AF65.

9 interact with U1-70K and U2AF35 but not with U2AF65.

10 to FACT and the splicing factors SF2/ASF and U2AF65.

11 the large subunit of the U2 auxiliary factor U2AF65.

12 of ATXN1 with the splicing factors RBM17 and U2AF65.

13 s for 14-3-3, and splicing factors RBM17 and U2AF65.

14 cleoprotein auxiliary factor 65-kDa subunit (U2AF65).

15 splicing through interfering with binding of U2AF(65).

16 ribute to degenerate Py-tract recognition by U2AF(65).

17 for universal 3' splice site recognition by U2AF(65).

18 nucleoplasm, and RGH3alpha colocalizes with U2AF(65).

19 h the 65-kDa subunit of U2 auxiliary factor (U2AF65), a protein associated with the 3' splice site.

20 he exogenously expressed large U2AF subunit, U2AF65, accumulates in spliced mRNP, leading to the recr

21 em RNA recognition motif domains (RRM1,2) of U2AF65 adopt closed/inactive and open/active conformatio

22 The D231V amino acid change restores U2AF(65) affinity for two mutated splice sites that caus

23 the 5' regions of Py tracts severely reduce U2AF(65) affinity.

24 proteins and SR related proteins, including U2AF65, all of which are known to function in alternativ

25 Coexpression of a plant orthologue of U2AF65 alleviated the splicing repression of AtPTB1.

26 We showed that U2AF(65) and Aly, two proteins known to interact with UA

27 We show that U2AF(26) can associate with U2AF(65) and can functionally substitute for U2AF(35) in

28 ly identified, UHM/ULM-mediated complexes of U2AF(65) and SPF45 with SF3b155, this work demonstrates

29 SF1 phosphorylation on its associations with U2AF(65) and splice-site RNAs are likely to influence pr

30 Specific hydrogen bonds between U2AF(65) and the uracil bases provide an explanation for

31 e RS domain of the essential splicing factor U2AF(65) and then in the prespliceosome by the ESE bound

32 r class of introns as a ternary complex with U2AF(65) and U2AF(35) splicing factors.

33 The large (U2AF65) and small (U2AF35) subunits of the U2AF heterodi

34 e factor U2 auxiliary factor 65 kDa subunit (U2AF65) and to act as a modulator of alternative splicin

35 of complex A is associated with the loss of U2AF65 and 35.

36 licing enhancers and their binding proteins (U2AF65 and ASF/SF2) that had critical roles in splicing

37 e ESS showed that the U-rich 5' region binds U2AF65 and polypyrimidine tract binding protein, the C-r

38 2AF35 directly mediates interactions between U2AF65 and proteins bound to the enhancers.

39 We provide evidence that both U2AF65 and PRP19C are required for CTD-dependent splicin

40 for CTD-dependent splicing activation, that U2AF65 and PRP19C interact both in vitro and in vivo, an

41 eraction results in increased recruitment of U2AF65 and PRP19C to the pre-mRNA.

42 s with the 3' splice site recognition factor U2AF65 and recruits it to the p21(Cip1) gene and mRNA.

43 mplex containing RBM39, which interacts with U2AF65 and SF3b155 and promotes U2 snRNP recruitment to

44 d with DDS enhanced both -KTS WT1 binding to U2AF65 and splicing-factor colocalization.

45 of Ataxin-1 with the spliceosome-associated U2AF65 and the adaptor 14-3-3 proteins.

46 Thus, U2AF35 functions as a bridge between U2AF65 and the enhancer complex to recruit U2AF65 to the

47 complex of two well-known splicing factors: U2AF65 and the Prp19 complex (PRP19C).

48 nd dynamics of 3' splice site recognition by U2AF65 and the role of U2AF35 in the U2AF heterodimer, w

49 icated in selection, the proteins U2AF35 and U2AF65 and the U2 snRNP, are able to recognize alternati

50 a heterodimer with subunits of 65 and 35 kD (U2AF65 and U2AF35).

51 pecific, sequential recognition by U1 snRNP, U2AF65 and U2AF35.

52 riptomes of HEK293 cells with varying U2AF35/U2AF65 and U2AF35a/b ratios.

53 Mammalian U2AF65 and UAP56 are required for prespliceosome (PS) fo

54 factor) is a heterodimer composed of 65-kDa (U2AF(65)) and 35-kDa (U2AF(35)) subunits.

55 is a heterodimer comprising a large subunit, U2AF65, and a small subunit, U2AF35.

56 interacts with an essential splicing factor, U2AF65, and associates with the splicing machinery.

57 ch is recruited to the pre-mRNA dependent on U2AF65, and is required for the U2 snRNP-branchpoint int

58 show that SF1 interacts strongly with human U2AF65, and that SF1 is a bona fide E complex component.

59 Furthermore, MBNL1 and U2AF65 appear to compete by binding to mutually exclusiv

60 intron to bind to a splicing factor such as U2AF65, as determined by an RNA electrophoretic mobility

61 use this information to clone a human 56-kD U2AF65 associated protein (UAP56).

62 The human 56-kD U2AF(65)-associated protein (hUAP56), a member of the DE

63 in hnRNP C competes with the splicing factor U2AF65 at many genuine and cryptic splice sites.

64 P is not required for splicing or loading of U2AF65 at other investigated p53-induced targets, includ

65 ressor hnRNP A1 and interfering with that of U2AF65 at the 3' splice site of exon 7.

66 osin, the consensus polypyrimidine tract for U2AF65, AUUUA repeats and r(U)20were used as competitors

67 rylation reduces phospho-SF1 and phospho-SF1-U2AF(65) binding affinities for either optimal or subopt

68 hile maintaining its uridine identity blocks U2AF(65) binding and splicing.

69 We conclude that reduced U2AF(65) binding is a molecular consequence of disease-r

70 ecifically and regulate splicing by blocking U2AF(65) binding to the 3' splice site upstream of exon

71 tions that strengthen the stem-loop decrease U2AF65 binding affinity and also repress exon 5 inclusio

72 s RNA nonspecifically and that the sites for U2AF65 binding and RNA binding are overlapping (or the s

73 ers but may be partially attributed to lower U2AF65 binding and weaker splicing-related motifs.

74 Thus, U2AF65 binding can be blocked either by MBNL1 binding or

75 a remarkably distinct manner from the local U2AF65 binding mode of SF3b155.

76 RNA oligonucleotide containing the consensus U2AF65 binding site, but U2AF65 was not displaced by a n

77 synthesizing a transcript segment that is a U2AF65 binding site.

78 The multiple U2AF65 binding sites on SF3b155 suggest a model in which

79 terminal domain of SF3b155 contains multiple U2AF65 binding sites that are distinct from the binding

80 We propose that the multiple U2AF65 binding sites within SF3b155 regulate conformatio

81 Thus, by preventing U2AF65 binding to Alu elements, hnRNP C plays a critical

82 vitro splicing, we found it had no effect on U2AF65 binding.

83 1 sequences are required for MBNL1-activated U2AF65 binding.

84 e similarity to the previously characterized U2AF65-binding domain of SF1.

85 relative molecular mass (Mr 65K) of 65,000 (U2AF65) binds to the poly(Y) tract, whereas the role of

86 metic aspartate and to alanine, we show that U2AF65 binds Ataxin-1 in a Ser776 phosphorylation indepe

87 Providing the link to the CTD, we show that U2AF65 binds directly to the phosphorylated CTD, and tha

88 We show that Neurospora U2AF65 binds RNA with low affinity and specificity, appa

89 U2AF65 binds specifically to 3' splice sites, but previo

90 ortion of the intron as a stem-loop, whereas U2AF65 binds the same region in a single-strand structur

91 U2AF65 binds to RNA at the polypyrimidine tract, whereas

92 mini-gene, and the results demonstrate that U2AF65 binds to such a site and controls the mRNA stabil

93 The mammalian splicing factor U2AF65 binds to the polypyrimidine tract adjacent to the

94 Early in mammalian spliceosome assembly, U2AF65 binds to the pyrimidine tract between the BPS and

95 e determined six crystal structures of human U2AF(65) bound to cytidine-containing Py tracts.

96 e also determined two baseline structures of U2AF(65) bound to the deoxy-uridine counterparts and com

97 cooperative interaction with splicing factor U2AF65 bound to an adjacent polypyrimidine tract (PPT) f

98 In the current model, U2AF65, bound at the polypyrimidine tract of the intron,

99 Arabidopsis AtU2AF65b encodes a putative U2AF65 but its specific functions in plants are unknown.

100 point binding protein (BBP) (SF1) and Mud2p (U2AF65), but lacks an obvious U2AF35 homolog, leaving op

101 thought to play a role in the recruitment of U2AF(65) by serine-arginine-rich (SR) proteins in enhanc

102 n JMJD6 and U2AF65, revealing that JMJD6 and U2AF65 co-regulated a large number of alternative splici

103 rative 3' splice site recognition by the SF1-U2AF(65) complex (where cooperativity is defined as a no

104 2AF(65) splicing factors, as well as the SF1/U2AF(65) complex in the absence and presence of AdML (ad

105 n the maximum dimension, relative to the SF1/U2AF(65) complex in the absence of RNA ligand.

106 peptides demonstrates that formation the SF1/U2AF65 complex is likely to affect regions of SF1 beyond

107 t or extended inter-RRM proximities from the U2AF(65) conformational pool.

108 The majority of U2AF(65) conformations exhibit few contacts between the

109 RRM spacings from a pre-existing ensemble of U2AF(65) conformations.

110 The spatial distribution of U2AF65 conformations is found to be highly anisotropic,

111 U2AF65 contains an RNA binding domain, required for inte

112 cing factor 1 (SF1) and U2 auxiliary factor (U2AF(65)) cooperatively recognize the 3' splice site dur

113 , and JMJD6-mediated lysine hydroxylation of U2AF65 could account for, at least partially, their co-r

114 he RS domain from a related splicing factor, U2AF65, could not rescue viability and was inactive in i

115 , particularly to the Arg-Ser (RS) domain of U2AF65, creates Tat inhibitors that localize to subnucle

116 Here we show that U2AF65 crosslinking is replaced by crosslinking of three

117 We found that the crystal structure of the U2AF(65)-D231V variant confirms favorable packing betwee

118 ed from the high resolution coordinates of a U2AF65 deletion variant bound to RNA.

119 The isoform-specific U2AF35 expression was U2AF65-dependent, required interactions between the U2AF

120 U2AF65 directly contacts the polypyrimidine tract and is

121 Here, we report that two members of the U2AF65 family of proteins, hCC1.3, which we call CAPERal

122 y, we identified a preferred binding site of U2AF(65) for purine substitutions in the 3' regions of P

123 cities of wild-type and site-directed mutant U2AF(65) for region-dependent cytosine- and uracil-conta

124 directly with the essential splicing factor U2AF65 for binding at the 3' end of intron 4.

125 Despite the central importance of U2AF65 for splice site recognition, the relative arrange

126 ate the essential nature of the third RBD of U2AF65 for the interaction between the two proteins, bot

127 pen reading frame is similar to that for the U2AF65 from mammals and flies.

128 A U2AF65 fusion named T-RS interacts with the nonphosphory

129 The essential pre-mRNA splicing factor, U2AF(65), guides the early stages of splice site choice

130 The splicing and nuclear export factor U2AF65 has the opposite effect, decreasing expHTT nuclea

131 central domain and a proline-rich region of U2AF65, has been determined at 2.2 A resolution.

132 Moreover, the fission yeast U2AF65 homolog, Prp2/Mis11 protein, was phosphorylated m

133 pre-mRNA splicing factor U2AF2 (also called U2AF65) identifies polypyrimidine (Py) tract signals of

134 ating a substrate that recruits U1 snRNP and U2AF65 in a splice signal-independent manner.

135 gs reveal an intimate link between JMJD6 and U2AF65 in alternative splicing regulation, which has imp

136 Overexpression of U2AF65 in cells transfected with a PLE-containing report

137 protein SC35 can functionally substitute for U2AF65 in the reconstitution of pre-mRNA splicing in U2A

138 o acids, KTS, show stronger interaction with U2AF65 in vitro and better colocalization with splicing

139 ng of the normally intronic splicing factor, U2AF65, inhibits splicing.

140 splicing assays reveal unforeseen roles for U2AF(65) inter-domain residues in recognizing a contiguo

141 Furthermore, the complex of the SF1/U2AF65 interacting domains is stabilized by 3.3 kcal mol

142 mol-1 relative to the complex of the SF3b155/U2AF65 interacting domains, consistent with the need for

143 this paper we demonstrate that the mBBP/SF1-U2AF65 interaction promotes cooperative binding to a bra

144 ifts and sequence requirements indicate that U2AF65 interactions with each of the SF3b155 sites are s

145 U2AF(65) is essential for pre-mRNA splicing in most euka

146 The essential pre-mRNA splicing factor U2AF(65) is faced with the paradoxical tasks of accurate

147 ively poorly conserved in higher eukaryotes, U2AF(65) is faced with the problem of specifying uridine

148 The U2 auxiliary factor large subunit (U2AF65) is an essential pre-mRNA splicing factor for the

149 In contrast, U2AF65 is more loosely bound in C28 transcription comple

150 When U2AF65 is prevented from binding to the pre-mRNA, the U2

151 NA is still contained within the polymerase, U2AF65 is tightly bound.

152 s RNA may represent an adaption for reducing U2AF65 levels when pre-mRNA levels are low.

153 The U2AF(65) linker residues between the dual RNA recognitio

154 ins of SR- and SR-related proteins including U2AF65, Luc7-like protein 3 (Luc7L3), SRSF11 and Acinus

155 with transcription regulator BRD4, histones, U2AF65, Luc7L3, and SRSF11.

156 on SF3b155 suggest a model in which multiple U2AF65 molecules bound to the intron could enhance U2 sn

157 In mammalians and yeast, the splicing factor U2AF65/Mud2p functions in precursor messenger RNA (pre-m

158 RNAs are often interrupted with purines, yet U2AF(65) must identify these degenerate Py-tracts for ac

159 The essential splicing factor U2AF(65) normally recognizes a Py tract consensus sequen

160 cleoprotein auxiliary factor 65 kDa subunit (U2AF65) on the upstream intron.

161 ation is sufficient to explain the action of U2AF65 on spliceosome components located both 5' and 3'

162 This suggests that MBNL1 and U2AF65 play a role in nuclear export of expHTT RNA.

163 NA recognition motif (RRM) of U2AF35 and the U2AF65 polyproline segment interact via reciprocal "tong

164 ecific binding of phospho-SF1 to its cognate U2AF(65) protein partner.

165 Addition of U2AF65 protein further enhanced the splicing reconstitut

166 ablished by analysis of site-directed mutant U2AF(65) proteins using surface plasmon resonance.

167 several SR proteins but also with U1-70K and U2AF65, proteins associated with 5' and 3' splice sites,

168 arge enthalpy-entropy compensation underlies U2AF65 recognition of an optimal polyuridine tract.

169 consecutive RNA recognition motifs (RRM) of U2AF(65) recognize a polypyrimidine tract at the 3' spli

170 Tandem RNA recognition motifs (RRM)s of U2AF65 recognize polypyrimidine tract signals adjacent t

171 To understand how U2AF(65) recognizes degenerate Py tracts, we determined

172 How the essential pre-mRNA splicing factor U2AF(65) recognizes the polypyrimidine (Py) signals of t

173 Since U2AF65 recognizes the polypyrimidine tract (PPT) and for

174 A splicing factor, U2 auxiliary factor 65KD (U2AF(65)) recognizes the polypyrimidine tract (Py-tract)

175 rich domain at the N-terminus of Neurospora U2AF65 regulates its RNA binding.

176 nd show that their preferential responses to U2AF65-related proteins and SRSF3 are associated with un

177 In this report we identify a new region of U2AF65 required for function, and use this information t

178 ntaining proteins such as SC35, ASF/SF2, and U2AF65, restored the splicing activity of the Sip1-immun

179 RNA-dependent interaction between JMJD6 and U2AF65, revealing that JMJD6 and U2AF65 co-regulated a l

180 characteristics, the X-ray structure of the U2AF(65) RNA binding domain bound to a Py tract composed

181 rwise distance distribution functions of the U2AF65 RNA binding domain and those either previously ob

182 Here, the solution conformation of the U2AF65 RNA binding domain determined using small angle x

183 shapes and RNA interactions of the wild-type U2AF65 RNA binding domain were compared with those of U2

184 We determined four structures of an extended U2AF(65)-RNA-binding domain bound to Py-tract oligonucle

185 nges that are induced by assembly of the SF1/U2AF(65)/RNA complex serve to position the pre-mRNA spli

186 The molecular dimensions of the SF1/U2AF(65)/RNA complex substantially contracted by 15 A in

187 e of U2AF35, whose UHM domain interacts with U2AF65 RRM1, increases the population of the open arrang

188 es the population of the open arrangement of U2AF65 RRM1,2 in the absence and presence of a weak Py-t

189 ay structure of the N-terminal RRM domain of U2AF(65) (RRM1) is described at 1.47 A resolution in the

190 formational selection and induced fit of the U2AF(65) RRMs are complementary mechanisms for Py-tract

191 ay scattering to demonstrate that the tandem U2AF(65) RRMs exhibit a broad range of conformations in

192 recognition, the relative arrangement of the U2AF65 RRMs and the energetic forces driving polypyrimid

193 Thus, the U2AF65 RS domain plays a direct role in modulating splic

194 d found that the two Pys are responsible for U2AF65's binding with intron 4.

195 Notably, RNA-binding by U2AF(65) selectively stabilizes pre-existing alternative

196 RNAi silencing of the basal splicing factors U2AF65, SF1, and U2AF35.

197 These findings indicate that U2AF65 shifts from protein-protein to protein-RNA intera

198 , pre-mRNA for the essential splicing factor U2AF65 sometimes is spliced to produce an RNA that inclu

199 anges were observed for the isolated SF1 and U2AF(65) splicing factors or their individual complexes

200 and ab initio shape restorations of SF1 and U2AF(65) splicing factors, as well as the SF1/U2AF(65) c

201 Similarly to TAP, U2AF65 stimulated directly the nuclear export and expres

202 In contrast, a high-affinity site for U2AF65 strongly inhibited exon inclusion.

203 cts and potentially could rearrange when the U2AF(65) structure adapts to different Py tract sequence

204 Based on a comparison of new U2AF(65) structures bound to either A- or G-containing P

205 ted the interaction between ZC3H14 and U2AF2/U2AF(65) Taking all the findings into consideration, we

206 he complete spectrum of the unbound forms of U2AF65 that coexist with the small percentage of a prefo

207 evealed that Urp specifically interacts with U2AF65 through a U2AF35-homologous region and with SR pr

208 ur work demonstrates that in vivo binding of U2AF(65) to a polypyrimidine tract requires a flexible R

209 nformations may contribute to the ability of U2AF(65) to recognize a variety of Py-tract sequences.

210 the splicing defect is due to the failure of U2AF(65) to recognize the pseudouridylated polypyrimidin

211 AF(26) functions by enhancing the binding of U2AF(65) to weak 3' splice sites.

212 ell growth, AR-V7 levels, and recruitment of U2AF65 to AR pre-mRNA.

213 domain mutants indicated that the ability of U2AF65 to contact the branch point, to promote the U2 sn

214 est that MBNL1 directly activates binding of U2AF65 to enhance upstream intron removal to ultimately

215 lation shift toward the open conformation of U2AF65 to facilitate the recognition of weak Py-tracts a

216 Cross-linking of U2AF65 to RNA in a C28 complex was eliminated by the add

217 n U2AF65 and the enhancer complex to recruit U2AF65 to the adjacent intron.

218 lymerase II may function not only to deliver U2AF65 to the nascent transcript but also to modulate ef

219 Here it is reported that binding of U2AF65 to the polypyrimidine tract directed the RS domai

220 ex assembly by competing with the binding of U2AF65 to the polypyrimidine tracts of repressed 3' spli

221 cally reduced by the addition of recombinant U2AF65 to the transcription reaction.

222 rmational plasticity as a possible means for U2AF65 to universally identify diverse pre-mRNA splice s

223 posal is that enhancers stabilize binding of U2AF65 to weak polypyrimidine (Py) tracts.

224 Our results reveal a new function of U2AF65: to position a DEAD box protein required for U2 s

225 sociated with spliceosome formation (U2AF35, U2AF65, U1A, and U1-70K) correlated with splicing effici

226 Our results suggest that the U2AF65-U2AF35 complex identifies the U4CAG/R, with U2AF3

227 he branchpoint binding protein, SF1, and the U2AF65-U2AF35 heterodimer bound at an adjacent sequence.

228 etion showed that Urp is associated with the U2AF65/U2AF35 heterodimer.

229 rometry were used to show that rather than a U2AF65/U2AF35-like heterodimer, Mud2p forms a complex wi

230 icing requires a heterodimer of the proteins U2AF65/U2AF35.

231 K)nXRW(DE) consensus sequence for predicting U2AF65-UHM ligands from genomic sequences, where parenth

232 ntaining SF3b155 sites are recognized by the U2AF65-UHM using intrinsic tryptophan fluorescence exper

233 tein auxiliary factor 65-kilodalton subunit (U2AF65) undergoes posttranslational lysyl-5-hydroxylatio

234 e sampled by the multidomain splicing factor U2AF65 using complementary nuclear magnetic resonance sp

235 Previously, the structure of a U2AF(65) variant in complex with poly(U) RNA suggested t

236 evant mutations, and that a structure-guided U2AF(65) variant is capable of manipulating gene express

237 A binding domain were compared with those of U2AF65 variants containing either Py tract-binding prote

238 the SR domains of the SR proteins SRP75 and U2AF65, via fusion to a heterologous MS2 RNA binding pro

239 ining the consensus U2AF65 binding site, but U2AF65 was not displaced by a nonconsensus RNA.

240 d with decreased cross-linking of the factor U2AF65, whereas regulation at step 2 is correlated with

241 ulated binding can be recapitulated in human U2AF65 which has been mutated to decrease both affinity

242 e subunit of the U2 snRNP associated factor (U2AF65), which is essential for splicing of human intron

243 h the well studied mammalian splicing factor U2AF65, which binds to the adjacent polypyrimidine (PY)

244 Furthermore, overexpression of the truncated U2AF65, which contains the arginine and serine dipeptide

245 a Serine-Arginine-Rich (RS) domain-deficient U2AF65, which is defective in CDCA5 pre-mRNA processing.

246 ion and interaction with the splicing factor U2AF65, which promotes mRNA processing and the stabiliza

247 e spliceosome components, such as U2AF35 and U2AF65, whose splicing functions are modulated by PARP1-

248 f the U2 small nuclear RNA auxiliary factor (U2AF65) with the splicing factor 1 (SF1) or the spliceos

249 Therefore, the association of U2AF65 with RNA polymerase II may function not only to d

250 e scRNAPy interfered with the interaction of U2AF65 with the intron and repressed the IE2 expression.

251 , while the first Py enhanced the binding of U2AF65 with the intron.

252 interaction of the essential splicing factor U2AF65 with the pre-mRNA polypyrimidine tract.

253 dition of the small U2AF subunit (U2AF35) to U2AF65 with weakened RNA binding affinity significantly

254 nteraction between HCC1, a factor related to U2AF65, with both subunits of U2AF.

255 ed to find that a D231V amino acid change in U2AF(65) would specify U over other nucleotides.