ライフサイエンスコーパス: hnRNP A1

1 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1).

2 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1).

3 nctions by binding of the splicing repressor hnRNP A1.

4 tif that mediates bidirectional transport of hnRNP A1.

5 affinity and specificity similar to those of hnRNP A1.

6 les as the nucleocytoplasmic shuttle protein hnRNP A1.

7 ong similarity to the RNA-binding domains of hnRNP A1.

8 Importantly, tax MAbs reacted with hnRNP A1.

9 ponents, including the heteronuclear protein hnRNP A1.

10 of either RRM impairs splicing repression by hnRNP A1.

11 n and of the general pre-RNA-binding protein hnRNP A1.

12 gle-stranded overhangs of human telomeres by hnRNP A1.

13 ing site for the pre-mRNA processing protein hnRNP A1.

14 s conform to the consensus binding motifs of hnRNP A1.

15 nergistic manner with the RNA-binding factor hnRNP A1.

16 mediated by the cytoplasmic accumulation of hnRNP A1.

17 ent cells that express exogenous cytoplasmic hnRNP A1.

18 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), a protein with multiple roles in mRNA metabol

19 hTRN1 interacts with the M9 region of hnRNP A1, a 38-amino-acid domain rich in Gly, Ser, and A

20 We recently found that hnRNP A1, a protein implicated in many aspects of RNA pr

21 NP complexes from HeLa cells associated with hnRNP A1, a shuttling hnRNP protein.

22 icing of HPV18 E6E7 pre-mRNAs via binding to hnRNP A1, a well-characterized, abundantly and ubiquitou

23 A screen identified hnRNP A1 (A1) and RPS25 as IRES-binding trans-acting fac

24 creen to identify novel ITAFs and identified hnRNP A1 (A1) as a mediator of the IL-6 effect.

25 The RNA-binding protein hnRNP A1 (A1) was also critical to IL-6-stimulated trans

26 Telomere bound hnRNPs include hnRNP A1, A2-B1, D and E and telomerase bound hnRNPs inc

27 Most importantly, the expression of hnRNP A1/A2 and PTB/nPTB is significantly altered in pat

28 /CG9983, the fruit fly ortholog of the human hnRNP A1/A2 factors.

29 protein overexpression, and found two tandem hnRNP A1/A2 motifs within the ISS that are responsible f

30 essing cell extracts with an antibody to the hnRNP A1/A2 proteins, which are splicing and nuclear shu

31 ding proteins, such as hnRNP L, PTB/nPTB and hnRNP A1/A2.

32 inhibitory effect of ISTL1 is independent of hnRNP A1/A2B1 and PTB1 previously implicated in SMN2 exo

33 Overexpression of hnRNP A1 accelerated the kinetics of viral RNA synthesis

34 In the present study, we demonstrate that hnRNP A1 also participates in the transcription and repl

35 Importantly, knockdown of hnRNP A1 also renders quiescent Akt-containing cells sen

36 Mutants in the 5' INS that prevent hnRNP A1 and 50 kDa protein binding are inactive in the

37 nstrate that the related splicing repressors hnRNP A1 and A2, as well as the polypyrimidine-tract-bin

38 We used RNA sequencing (RNA-seq) to confirm hnRNP A1 and A2/B1 motif-dependent roles genome-wide, pr

39 a marked increase in the protein levels FUS, hnRNP A1 and hnRNP E2, three RNA-binding proteins involv

40 Although hnRNP A1 and HuR can both be immunoprecipitated from rib

41 voring recruitment of the splicing repressor hnRNP A1 and interfering with that of U2AF65 at the 3' s

42 sults reveal general rules of specificity of hnRNP A1 and provide a quantitative framework for unders

43 Thus, hnRNP A1 and PTB use different mechanisms to repress c-s

44 Furthermore, hnRNP A1 and PTB, both of which also bind to the complem

45 progression and a coordinated regulation by hnRNP A1 and Sam68 as modifiers of hypoxia-induced skipp

46 a treatment in cell culture led to increased hnRNP A1 and Sam68 levels.

47 ing of SMN exon 7 and was found to bind both hnRNP A1 and Sam68.

48 The activities of hnRNP A1 and SF2/ASF on N1 splicing were examined by add

49 n-proximal 5' splice sites, and tethering of hnRNP A1 and SF2/ASF proteins between competing splice s

50 Mutations in hnRNP A1 and SF2/ASF show that the opposite effects of t

51 differentiation, HPV16 infection upregulates hnRNP A1 and SF2/ASF, both key factors in alternative sp

52 ionship between the potential for binding of hnRNP A1 and U1 snRNA and the effect of hnRNP L on splic

53 heterogeneous nuclear ribonucleoprotein-A1 (hnRNP-A1), and a previously uncharacterized protein, FLJ

54 s nuclear ribonucleoprotein family (hnRNP L, hnRNP A1, and hnRNP M) were identified.

55 tax MAbs inhibited antibody binding to tax, hnRNP A1, and neurons.

56 ugh interaction with a host splicing factor, hnRNP A1, and regulates E6 and E7 expression of the earl

57 stitutive transcriptional repressor protein, hnRNP A1, and the decreased binding activity of hnRNP A1

58 d the presence of two mRNA-binding proteins, hnRNP A1, and the elav-like protein, HuR, both of which

59 HnRNP A1 antagonizes ASF/SF2 by promoting the distal cho

60 osition of betaARB (HuR alone versus HuR and hnRNP A1) appears to be dependent on the mRNA probe used

61 neous nuclear ribonucleoprotein (hnRNP) L or hnRNP A1 are Akt substrates during Treg induction and ha

62 The two RRMs of hnRNP A1 are closely related but have distinct functions

63 These data point to hnRNP A1 as a critical regulator of c-myc translation an

64 Although the importance of hnRNP A1 as a regulator of nuclear pre-mRNA and mRNA pro

65 Here we identify hnRNP A1 as an IRES trans-acting factor that regulates c

66 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) as the autoantigen.

67 eterogeneous nuclear ribonuclear protein-A1 (hnRNP-A1) as the autoantigen.

68 AKT phosphorylation of hnRNP A1 at serine 199 has been demonstrated to inhibit

69 NA, they facilitate cooperative spreading of hnRNP A1 between the two sites.

70 thus show that two cellular hnRNPs, PTB and hnRNP A1, bind to the transcription-regulatory sequences

71 We report here that hnRNP A1 binding and splicing repression can occur on an

72 restore exon 7 inclusion in vivo and prevent hnRNP A1 binding in vitro.

73 ng and in vitro evolution identify consensus hnRNP A1 binding motifs; however, such data do not revea

74 negative strand of the 3'-UTR also inhibited hnRNP A1 binding on the positive strand, indicating a po

75 ructed MHV mutants with a very high-affinity hnRNP A1 binding site inserted in place of, or adjacent

76 When the high affinity hnRNP A1 binding site was inserted into the beta-globin

77 This inserted hnRNP A1 binding site was not able to functionally repla

78 se change does indeed create a high-affinity hnRNP A1 binding site, and base substitutions that disru

79 In contrast, the mutant hnRNP A1 binding site, or binding sites for hnRNP C and

80 , potentially creates a second SMN2-specific hnRNP A1 binding site.

81 We show that the SC35 and the hnRNP A1 binding sites overlap within the juxtaposed ESE

82 -related exon-skipping mutations that create hnRNP A1 binding sites, but show that splicing can be re

83 Initial hnRNP A1 binding to an exonic splicing silencer at the 3

84 solution as a dimer and supports a model for hnRNP A1 binding to nucleic acids in arrays that may mak

85 We propose that hnRNP A1 binding to the ESS inhibits splicing of the ups

86 arose by cytoplasmic hnRNP A1, while nuclear hnRNP A1 binding was unaffected.

87 solution, which could potentially influence hnRNP A1 binding.

88 A high-affinity hnRNP A1-binding sequence can substitute functionally fo

89 ulatory elements as follows: a high affinity hnRNP A1-binding site (WA1), a sequence involved in Rev-

90 ective-interfering RNAs containing a mutated hnRNP A1-binding site have reduced RNA transcription and

91 Introduction of point mutations into the hnRNP A1-binding site or knockdown of hnRNP A1 expressio

92 A strong hnRNP A1-binding site was located 90 to 170 nucleotides

93 Mutation of hnRNP A1-binding sites prevented hypoxia-induced skippin

94 hnRNP A1 binds both IRESs in vitro and in intact cells a

95 hnRNP A1 binds to RNA in a cooperative manner.

96 Serine 199-phosphorylated hnRNP A1 binds to the IRESs normally but is unable to su

97 We show that hnRNP A1 binds to the loop of pri-miR-18a and induces a

98 Recombinant hnRNP A1 bound to these two RNA regions in vitro in a se

99 hroleukemia cell line, CB3, does not express hnRNP A1 but still supports MHV replication, suggesting

100 ing led us to test more directly the role of hnRNP A1 by analysis of MHV replication and RNA synthesi

101 progenitor 32Dcl3 cells, BCR/ABL stabilizes hnRNP A1 by preventing its ubiquitin/proteasome-dependen

102 We further show that hnRNP A1 can also spread in a 5'-to-3' direction, althou

103 hnRNP A1 can be proteolyzed to unwinding protein (UP1),

104 n addition, we show by NMR that both RRMs of hnRNP A1 can bind simultaneously to a single bipartite m

105 Moreover, hnRNP A1 can effectively unwind an RNA hairpin upon bind

106 ss-linking experiments show that SF2/ASF and hnRNP A1 compete to bind pre-mRNA, and we conclude that

107 To confirm that the hnRNP A1 complex is responsible for INS activity, a synt

108 e of UP1, the amino-terminal domain of human hnRNP A1 containing two RNA-recognition motifs (RRMs), b

109 A1 and depends on the formation of a FUS-Jun-hnRNP A1-containing complex and on lack of PKCbetaII pho

110 Nuclear and cytoplasmic hnRNP A1 could be distinguished by the differential sens

111 Antibodies to hnRNP-A1 cross-reacted with HTLV-1-tax, the immune respo

112 romoting distal splicing was also seen in an hnRNP A1-deficient MEL cell line.

113 tion on SMN2 exon 7 splicing is specific, as hnRNP A1 depletion has little or no effect on other inef

114 We demonstrate that the striking effect of hnRNP A1 depletion on SMN2 exon 7 splicing is specific,

115 n be restored only modestly or not at all by hnRNP A1 depletion.

116 With longer flap substrates the hnRNP A1 effect is more modest and is suppressed by the

117 Thus, cytoplasmic and nuclear hnRNP A1 exhibit different RNA binding profiles, perhaps

118 to the hnRNP A1-binding site or knockdown of hnRNP A1 expression promoted 233^416 splicing and reduce

119 The increase in AUBP activity of cytoplasmic hnRNP A1 following RNA polymerase II inhibition correlat

120 We determined the affinity of hnRNP A1 for all possible sequence variants (n = 16,384)

121 organization of the two RRMs is essential to hnRNP A1 function.

122 The cellular absence of hnRNP A1 had no detectable effect on the production of i

123 Whereas nuclear hnRNP A1 has been shown to bind intronic sequences and m

124 heterogeneous nuclear ribonucleoprotein A1 (hnRNP-A1) has been implicated in telomere protection and

125 Drosophila Hrp38, a homolog of human hnRNP A1, has been shown to regulate splicing, but its f

126 geneous nuclear ribonucleoproteins (hnRNPs), hnRNP A1, hnRNP A2/B1, and hnRNP Q, bind to the dengue v

127 The sumoylation of hnRNP A1, hnRNP F, and hnRNP K were confirmed in vivo by

128 This sequence was also specifically bound by hnRNP A1, hnRNP H, ASF/SF2 and SRp40, but not by 9G8.

129 s binding to this element were identified as hnRNP A1, hnRNP H, hnRNP F, and SF2/ASF by site-specific

130 Here, we identify the mRNAs bound to the hnRNP-A1, hnRNP-E2, hnRNP-K, and La/SSB RBPs in BCR/ABLt

131 vary gland cells with fluorescent hrp36, the hnRNP A1 homolog, and the nuclear envelope by fluorescen

132 Drosophila hnRNP A1, Hrp38, is required for germ line stem cell mai

133 a suggest that seemingly disparate roles for hnRNP A1 in alternative splice site selection, RNA proce

134 tional studies failed to indicate a role for hnRNP A1 in ARE-dependent mRNA turnover.

135 quired for the subcellular redistribution of hnRNP A1 in cells subjected to OSM.

136 esting that alternative proteins can replace hnRNP A1 in cellular functions and viral infection.

137 RNA foci with SRSF2, hnRNP H1/F, ALYREF and hnRNP A1 in cerebellar granule cells and with SRSF2, hnR

138 , stress-induced cytoplasmic accumulation of hnRNP A1 in MDA-MB-468 cells dynamically alters the CEAC

139 These results support a role for hnRNP A1 in mediating rapamycin-induced alterations of c

140 r findings demonstrate that the functions of hnRNP A1 in MHV RNA synthesis can be replaced by other c

141 involved in the cytoplasmic accumulation of hnRNP A1 in NIH 3T3 cells subjected to OSM.

142 This effect of hnRNP A1 in promoting distal splicing was also seen in a

143 hnRNPs have previously been shown to replace hnRNP A1 in regulating RNA splicing.

144 ha phosphorylation-dependent accumulation of hnRNP A1 in SGs.

145 gardless of the presence of P-TEFb, HEXIM or hnRNP A1 in the complex.

146 ay, which in turn results in accumulation of hnRNP A1 in the cytoplasm.

147 The exact role of HuR and hnRNP A1 in the regulation of beta-AR mRNA stability rem

148 cular mimicry between an infecting agent and hnRNP-A1 in autoimmune disease of the CNS.

149 The role of hnRNP-A1 in telomere protection also involves DNA-depend

150 Two of the human MARs bind hnRNP-A1 in vitro directly within a 35-bp sequence that

151 heterogeneous nuclear ribonucleoprotein A1 (hnRNP-A1) in vivo during transcriptional up-regulation o

152 Expressing GzmA-resistant hnRNP A1 inhibited GzmA-mediated cell death and rescued

153 ral CB3 cellular proteins similar in size to hnRNP A1 interacted with the MHV RNA.

154 y attenuates viral replication by abrogating hnRNP A1 interactions.

155 plicing towards the downstream site, whereas hnRNP A1 interferes with U1 snRNP binding such that 5'SS

156 These data suggest that hnRNP A1 is a cellular factor that regulates the RNA-dep

157 hnRNP A1 is a nucleocytoplasmic shuttling heterogeneous

158 Human hnRNP A1 is a versatile single-stranded nucleic acid-bin

159 We conclude that hnRNP A1 is able to direct unspliced globin pre-mRNA int

160 d rescued pre-mRNA splicing, suggesting that hnRNP A1 is an important GzmA substrate.

161 sequences and modulate splicing, cytoplasmic hnRNP A1 is associated with poly(A)+ RNA, indicating dif

162 Previous studies indicated that cytoplasmic hnRNP A1 is capable of high-affinity binding of reiterat

163 une response between HTLV-I tax and neuronal hnRNP A1 is contained within the human immunodominant ep

164 in testis out of six mouse tissues, whereas hnRNP A1 is down-regulated during germ cell development.

165 that the recruitment of cellular proteins by hnRNP A1 is important for MHV RNA synthesis.

166 sults suggest that the shuttling activity of hnRNP A1 is important for the nucleocytoplasmic traffick

167 We showed that hnRNP A1 is methylated by PRMT5 on two residues, R218 an

168 The stimulation by hnRNP A1 is most dramatic using DNA substrates with shor

169 These results strongly suggest that hnRNP A1 is not a required host factor for MHV discontin

170 The region consisting of residues 1-195 of hnRNP A1 is referred to as UP1.

171 Here we report that hnRNP-A1 is phosphorylated by DNA-PKcs during the G2 and

172 Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is a multipurpose RNA-binding protein (RBP) in

173 Heterogeneous ribonucleoprotein A1 (hnRNP A1) is a prototype for the family of eukaryotic RN

174 Heterogeneous ribonucleoprotein A1 (hnRNP A1) is an abundant nuclear protein that participat

175 Heterogeneous nuclear ribonucleoprotein (hnRNP A1) is involved in pre-mRNA splicing in the nucleu

176 The heterogeneous nuclear ribonucleoprotein, hnRNP A1, is an IRES transacting factor (ITAF) that regu

177 NP A1, and the decreased binding activity of hnRNP A1 leads to the overexpression of gamma chain in H

178 We show here that hnRNP A1 levels are increased in myeloid progenitor cell

179 trongly correlated with elevated Ser(P)(199)-hnRNP A1 levels in a panel of 22 glioblastomas.

180 documented changes in the abundance of these hnRNP A1-like proteins during embryogenesis.

181 Furthermore, as hnRNP A1 likely participates in mRNA export, it raises t

182 Although the Tap NLS and the hnRNP A1 M9 element are shown to compete for transportin

183 ally constitutively spliced, suggesting that hnRNP A1 may be a general inhibitor of proximal splicing

184 it is the major determinant of Tra2beta- and hnRNP A1-mediated regulation.

185 We propose that interactions between hnRNP A1 molecules bound to the exonic and intronic site

186 The hnRNP A1 mutant caused a global inhibition of viral mRNA

187 Similar to the wild-type hnRNP A1, the hnRNP A1 mutant complexed with an MHV polymerase gene pr

188 sion in the cytoplasm of a dominant-negative hnRNP A1 mutant that lacks the nuclear transport domain

189 ell-signaling pathways regulates the rate of hnRNP A1 nuclear import.

190 To assess the potential role of hnRNP A1 nucleocytoplasmic shuttling activity in normal

191 IRES activity by inducing phosphorylation of hnRNP A1 on serine 199.

192 The negative activity of hnRNP A1 on splicing was compared with that of PTB, a pr

193 Depletion of hnRNP A1 or L by RNAi in HEK-293 cells promoted exon 7 i

194 Reducing expression levels of hnRNP A1 or overexpressing a dominant negative version o

195 Consequently, in cells lacking hnRNP-A1 or DNA-PKcs-dependent hnRNP-A1 phosphorylation,

196 al gel and proteolysis studies, we establish hnRNP A1 (or structurally related proteins that are post

197 lation in vivo and correlates with increased hnRNP A1 phosphorylation.

198 our results indicate that DNA-PKcs-dependent hnRNP-A1 phosphorylation is critical for capping of the

199 he G2 and M phases and that DNA-PK-dependent hnRNP-A1 phosphorylation promotes the RPA-to-POT1 switch

200 cells lacking hnRNP-A1 or DNA-PKcs-dependent hnRNP-A1 phosphorylation, impairment of the RPA-to-POT1

201 ng was antagonized by cotransfection with an hnRNP A1 plasmid.

202 ecent evidence has further demonstrated that hnRNP-A1 plays a crucial role in maintaining newly repli

203 Previous studies suggested that hnRNP A1 preferentially binds (under nonequilibrium cond

204 We additionally demonstrate that cytoplasmic hnRNP A1 preferentially binds ARE relative to pre-mRNAs

205 RNA interference to knock down hnRNP A1 prevented an IL-6 increase in myc protein expre

206 As hnRNP A1 propagates toward the 5' end of the exon, it an

207 he Kap beta2-mediated nuclear import of host hnRNP A1 protein and, in this way, favors virion formati

208 novel 38 amino acid transport signal in the hnRNP A1 protein, termed M9, which confers bidirectional

209 These studies suggest that hnRNP A1-PTB interactions provide a molecular mechanism

210 With purified components, hnRNP A1 reduces U1 snRNP binding to 5'SSs by binding co

211 k, together with the discovery of E2F3 as an hnRNP-A1-regulated factor, outlines the relevant role pl

212 HnRNP A1 regulates many alternative splicing events by t

213 Here, we provide the mechanism by which hnRNP A1 regulates this event.

214 During MHV infection, hnRNP A1 relocalizes from the nucleus to the cytoplasm,

215 SF2/ASF and in agreement with other systems, hnRNP A1 repressed c-src splicing in vitro.

216 tide hairpin loop contains the high-affinity hnRNP-A1-responsive 5'-UAGU-3' element and a proximal 5'

217 differentiation, and knockdown of hnRNP L or hnRNP A1 results in the lower induction of Treg cells.

218 RNA immunoprecipitation of hnRNP L and hnRNP A1 revealed a binding motif located central and 3'

219 nformational change to assemble a functional hnRNP A1-RNA complex.

220 Mechanisms and putative hnRNP A1-RNA interactions have been inferred primarily f

221 esented here provide the first insights into hnRNP A1-RNA interactions.

222 ittle is known about the structural basis of hnRNP A1-RNA recognition.

223 esembles sequence elements of several native hnRNP A1-RNA stem loop targets.

224 Here we describe a phosphomimetic mutant of hnRNP A1 (S199E) that is capable of binding both the cyc

225 monstrate that the phosphorylation status of hnRNP A1 serine 199 regulates the AKT-dependent sensitiv

226 hnRNP A1 shuttles between the nucleus and cytoplasm and

227 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) shuttles between the cytoplasm and nucleus and

228 Moreover, interference with hnRNP A1 shuttling activity resulted in downmodulation o

229 ated in CML-BCCD34+ in a BCR/ABL kinase- and hnRNP-A1 shuttling-dependent manner.

230 rentially spliced gene isoforms in LIN28 and hnRNP A1 small interfering RNA (siRNA)-treated cells.

231 ence of Tra2beta, ASF/SF2 and SRp40, whereas hnRNP A1 specifically inhibits this choice.

232 The physical determinants of hnRNP A1 splice site recognition remain poorly defined i

233 d calorimetric and NMR titrations of several hnRNP A1 subdomains into ESS3.

234 Overexpression of hnRNP A1 suppressed basal gamma-fibrinogen transcription

235 loops represent an important class of known hnRNP A1 targets, yet little is known about the structur

236 ein export, even for proteins such as GR and hnRNP A1 that use distinct nuclear export pathways.

237 ) bound to one of its substrates, the NLS of hnRNP A1, that we report here explains the mechanism of

238 Similar to the wild-type hnRNP A1, the hnRNP A1 mutant complexed with an MHV poly

239 However, in contrast to the wild-type hnRNP A1, the mutant protein failed to bind a 250 kDa ce

240 the transcription factor E2F3 associates to hnRNP-A1 through a conserved binding site located in the

241 hnRNP A1 thus has been proposed as a host factor in MHV

242 hnRNP L represses CD45 exon 4 by recruiting hnRNP A1 to a sequence upstream of the 5' splice site.

243 into the basis for high-affinity binding of hnRNP A1 to certain RNA sequences, and for nucleic acid

244 ings are relevant to the specific ability of hnRNP A1 to serve distinct roles in post-transcriptional

245 nown exonic splicing regulators, SF2/ASF and hnRNP A1, to the splicing of an exon primarily controlle

246 This study shows that MARs recruit and bind hnRNP-A1 upon transcriptional up-regulation.

247 , a synthetic high-affinity binding site for hnRNP A1 was also analysed.

248 The importance of hnRNP A1 was demonstrated by induction of apoptosis in e

249 A similar effect of hnRNP A1 was demonstrated with mutant hamster adenine ph

250 RE-specific binding protein in cells lacking hnRNP A1 was purified from CB3 mouse erythroleukemia cel

251 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) was identified as a component of the complexes

252 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), which forms a direct complex with FEN-1 and s

253 by inducing the cytoplasmic accumulation of hnRNP A1, which attenuates internal ribosome entry site-

254 binding site for a known repressor protein, hnRNP A1, which binds to SMN2 but not SMN1 exon 7 RNA.

255 Furthermore, the amount of hnRNP A1, which has been implicated in the export of cel

256 We show here that hnRNP A1, which shifts splicing towards an upstream 5'SS

257 r ribonucleoproteins [hnRNP]) represented by hnRNP A1, which shuttle continuously between the nucleus

258 tivity) and poly(U)-Sepharose by cytoplasmic hnRNP A1, while nuclear hnRNP A1 binding was unaffected.

259 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), whose structure was previously solved at 1.75

260 s methylation facilitates the interaction of hnRNP A1 with IRES RNA to promote IRES-dependent transla

261 n the nucleocytoplasmic shuttling pathway of hnRNP A1 with its bound RNAs.

262 phosphorylation modulates the interaction of hnRNP A1 with transportin Trn1.