ライフサイエンスコーパス: NELF

1 NELF and DSIF act together to inhibit transcription elon

2 NELF and DSIF collaborate to inhibit elongation by RNA p

3 NELF antisense experiments indicate that a reduction in

4 NELF causes Pol II to pause in the promoter-proximal reg

5 NELF colocalizes with RNAP II, and its level increases f

6 NELF depletion also delayed the dissociation of HSF from

7 NELF interacts with Pcf11, a transcription termination f

8 NELF is expressed in PNS and CNS tissues during embryoni

9 NELF-associated pausing of Pol II might be an obligatory

10 NELF-B is a BRCA1-interacting protein and subunit (with

11 NELF-B is predicted to form a HEAT repeat fold, also bin

12 NELF-mediated stalling of RNAPII also attenuates transcr

13 addition, we identify NCoR1-GPS2-HDAC3 as a NELF-interacting corepressor complex that is associated

14 bition of transcription, we did not detect a NELF-RNA contact when the nascent transcript was between

15 We identify a NELF core subcomplex formed by conserved regions in subu

16 Using a new competition assay, NELF-A and NELF-B are each shown to act independently as competitiv

17 of GR-regulated transactivation, NELF-A and NELF-B, relative to other factors in the overall gene in

18 equired for full activity of both NELF-A and NELF-B.

19 by conserved regions in subunits NELF-A and NELF-C, and resolve its crystal structure.

20 d throughout their lengths, while NELF-A and NELF-E contain nonconserved regions inserted between con

21 mediated phosphorylation of Spt5, NELF-A and NELF-E results in the dissociation of NELF from Pol II,

22 1, whereas SEC recruits P-TEFb to NELF-A and NELF-E via Paf1c and Med26, respectively.

23 The amino acid sequences of NELF-B and NELF-D are highly conserved throughout their lengths, wh

24 s extensive colocalization of the NELF-B and NELF-D subunits at hundreds of interbands.

25 of mutagenized libraries of GFP-binding and NELF-E-binding aptamers to their respective targets and

26 ELF, reverse the negative effect of DSIF and NELF and simultaneously facilitate the action of TFIIF.

27 gation, including components of the DSIF and NELF complexes.

28 ough a capping-independent block of DSIF and NELF loading.

29 However, the mechanism by which DSIF and NELF participate in setting up the paused Pol II remains

30 Our results show that DSIF and NELF require a nascent transcript longer than 18 nt to s

31 EFb reverses the negative effect of DSIF and NELF through a mechanism dependent on its kinase activit

32 Although DSIF and NELF were both required for inhibition of transcription,

33 The association of DSIF and NELF with initiated RNA Polymerase II (Pol II) is the ge

34 at TFIIF functionally competes with DSIF and NELF, and this competition is dependent on the relative

35 g the elongation complex containing DSIF and NELF, reverse the negative effect of DSIF and NELF and s

36 mbryos containing reduced levels of DSIF and NELF.

37 l link between PARP-1, ADP-ribosylation, and NELF.

38 ay induced by negative factors Spt5/Spt4 and NELF, which is overcome by the positive factor P-TEFb (C

39 complex, along with pausing factors SPT5 and NELF-A, at the intragenic CTCF-cohesin binding sites.

40 p distinct from those controlled by Spt5 and NELF.

41 d that IFN-induced recruitment of P-TEFb and NELF/DSIF was under the control of BRD4.

42 on the relative concentrations of TFIIF and NELF.

43 single-stranded nucleic acids in vitro, and NELF-C associates with RNA in vivo.

44 COBRA1 (also known as NELF-B) is a BRCA1-binding protein that regulates RNA po

45 ive transcription elongation factors such as NELF, DSIF, and factor 2.

46 Using a new competition assay, NELF-A and NELF-B are each shown to act independently as

47 al for antiviral immunity in insects because NELF and P-TEFb are required to restrict viral replicati

48 4) show that upon activation, eRNAs can bind NELF and are necessary for its transient removal from pr

49 pposite face of the NELF-AC subcomplex binds NELF-B.

50 d that is required for full activity of both NELF-A and NELF-B.

51 s raises the possibility that RNA binding by NELF is not necessary in promoter-proximal pausing.

52 KSHV OriLytL-K7 lytic genes is inhibited by NELF during latency, but can also be promptly reactivate

53 results reveal an earlier stage, mediated by NELF, when repression occurs at the HIV LTR.

54 rget genes were, like Hsp70, up-regulated by NELF-depletion, whereas the majority of target genes sho

55 transcriptional activator, HSF, might cause NELF to dissociate from the elongation complex.

56 Cohesin, NELF, and Spt5 pausing and elongation factor knockdown e

57 and the negative elongation factor complex (NELF).

58 ymerase II (Pol II) and the pausing complex, NELF and DSIF, are detected near the transcription start

59 Consequently, NELF knockdown resulted in significant reduction in DNA

60 Decreasing NELF also correlated with displacement of a positioned n

61 Decreasing NELF expression overcomes RNAP II pausing to enhance HIV

62 Depleting NELF increased processive HIV transcription and replicat

63 hromatin immunoprecipitation analyses detect NELF at the promoters of the hsp70 and beta1-tubulin gen

64 Drosophila NELF has four subunits similar to subunits of human NELF

65 Here, further characterization of Drosophila NELF is provided.

66 We analyzed the interactions among DSIF, NELF, and a reconstituted Drosophila Pol II elongation c

67 DSIF/NELF inhibits early transcript elongation until it is co

68 nascent RNA, TFIIS inhibition may help DSIF/NELF negatively regulate productive transcription.

69 These two activities of DSIF/NELF appear to be mechanistically distinct.

70 rt a previously undescribed activity of DSIF/NELF, namely inhibition of the transcript cleavage facto

71 mics of interactions between HIV-1 Tat, DSIF/NELF, and the transcription complexes actively engaged i

72 Our results demonstrate that DSIF/NELF associates with RNA pol II complexes during early t

73 FP and negative elongation factor subunit E (NELF-E) proteins with their corresponding canonical and

74 A common motif in each NELF was identified that is required for full activity o

75 Knockdown of either NELF or DSIF results in an increase in the levels of uaR

76 egatively regulate transcription elongation, NELF, Spt5, and Pcf11.

77 Stable knockdown of endogenous NELF-B has the opposite effects on an exogenous gene.

78 Reduction of the endogenous NELF proteins in breast cancer cells using small interfe

79 ecruitment of the negative elongation factor NELF at start sites.

80 th the polymerase negative elongation factor NELF.

81 that includes the negative elongation factor NELF.

82 blation of the primary pause-inducing factor NELF does not increase expression of lineage markers, bu

83 rough knockdown of the pause-inducing factor NELF leads to broadly attenuated immune gene activation.

84 y the complex of negative elongation factor (NELF) and 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazo

85 Negative elongation factor (NELF) and 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazo

86 IF together with negative elongation factor (NELF) associates with RNA polymerase II during early elo

87 occupancy of the negative elongation factor (NELF) at the p21(CIP1) promoter, although the level of b

88 ed by the host's negative elongation factor (NELF) at the promoter regions of OriLytL-K7 lytic genes

89 t release of the negative elongation factor (NELF) complex and productive elongation.

90 a decoy for the negative elongation factor (NELF) complex upon induction of immediate early genes (I

91 -E) of the human negative elongation factor (NELF) complex, which participates in RNA polymerase II p

92 ed member of the negative elongation factor (NELF) complex.

93 e pause-inducing negative elongation factor (NELF) complex.

94 The principal negative elongation factor (NELF) contains four polypeptides and requires for activi

95 hanisms by which negative elongation factor (NELF) establishes and maintains HIV latency.

96 The role of the negative elongation factor (NELF) in maintaining HIV latency was investigated follow

97 Negative elongation factor (NELF) induces RNAP II promoter proximal pausing and limi

98 he negative transcription elongation factor (NELF) inhibits basal transcription from the long termina

99 The human negative elongation factor (NELF) is a four-subunit protein complex that inhibits th

100 The Negative Elongation Factor (NELF) is a transcription regulatory complex that induces

101 thway, including negative elongation factor (NELF) that pauses RNA polymerase II (Pol II) and positiv

102 ns, depletion of negative elongation factor (NELF), a key factor in setting up paused Pol II, reduced

103 tes and inhibits negative elongation factor (NELF), a protein complex that regulates promoter-proxima

104 actor (DSIF) and negative elongation factor (NELF), act as negative transcription elongation factors

105 nit of the human negative elongation factor (NELF), directly binds to ERalpha and represses ERalpha-m

106 actors (BRD4 and negative elongation factor (NELF)-E) and to define their sites and mechanisms of act

107 s (DSIF) and the negative elongation factor (NELF).

108 es the 4-subunit negative elongation factor (NELF).

109 deletion in the nasal embryonic LHRH factor (NELF) gene in pedigree 1 and an additional heterozygous

110 l factor termed nasal embryonic LHRH factor (NELF) that was discovered in a differential screen of mi

111 The pausing factors NELF and DSIF are associated with these antisense polyme

112 vity of TTF2, and influenced pausing factors NELF and DSIF, but did not affect the function of TFIIS

113 factors we describe are the pausing factors--NELF (negative elongation factor) and DSIF (DRB sensitiv

114 Following NELF-E knockdown or tumor necrosis factor alpha (TNF-alp

115 This work thus uncovers a key role for NELF-mediated pausing in establishing the responsiveness

116 r associates with 39% of the genes that have NELF.

117 discern the functional architecture of human NELF through X-ray crystallography, protein crosslinking

118 obtain a genome-wide understanding of human NELF-mediated transcriptional regulation in vivo, we car

119 s four subunits similar to subunits of human NELF.

120 ion of hsp70 results in a marked decrease in NELF at the hsp70 promoter.

121 nse experiments indicate that a reduction in NELF expression decreases olfactory axon outgrowth and t

122 of negative elongation factors that include NELF and DSIF.

123 mall interfering RNA knockdown of individual NELF subunits.

124 Indeed, ESCs lacking NELF have dramatically attenuated FGF/ERK activity, rend

125 Like NELF-associated genes, most BEAF-associated genes are hi

126 Likewise, NELF and DSIF prior to stimulation were hardly detectabl

127 ption initiation and by assisting in loading NELF onto Pol II after initiation.

128 ponents: GR < Cdk9 < BRD4 </= induced gene < NELF-E.

129 rticoid receptor (GR), reporter, TIF2, NCoR, NELF-A, sSMRT, and STAMP) using our recently developed c

130 In contrast, no NELF or polymerase is detectible near the IP-10 promoter

131 shock induction, DSIF and polymerase but not NELF were strongly recruited to chromosomal puffs harbor

132 wn of the transcription factor SPT5, but not NELF-E, also gives rise to a specific inhibition of HSV-

133 We now report new activities of NELF-B and other NELF complex subunits, which are to att

134 tion for genetic and biochemical analysis of NELF in Drosophila.

135 However, novel actions of BRD4 and of NELF-E in GR-controlled gene induction have been uncover

136 A shRNA-based knockdown assay of NELF revealed that it negatively regulates the passage o

137 est a diverse transcriptional consequence of NELF-mediated RNAPII pausing in the human genome.

138 microarray analysis of S2 cells depleted of NELF and discovered that NELF RNAi affects many rapidly

139 Instead, depletion of NELF delayed the time taken for these genes to shut off

140 Consequently, depletion of NELF expression induced transition of stalled RNAPII int

141 Upon depletion of NELF-A, -C, or -E, the vast majority of NELF-regulated g

142 -A and NELF-E results in the dissociation of NELF from Pol II, thereby transiting transcription from

143 highly activated by the combined effects of NELF-E depletion and activation of initiation by TNF-alp

144 A positively charged face of NELF-AC is involved in RNA binding, whereas the opposite

145 architecture and three RNA-binding faces of NELF.

146 ne, eiger, displayed all of the hallmarks of NELF-dependent polymerase stalling.

147 Immunodepletion of NELF also impairs promoter proximal pausing on the hsp70

148 Immunodepletion of NELF or DSIF from a nuclear extract desensitizes transcr

149 1 in gastric cancer cells was independent of NELF-E.

150 lated with and probably results from loss of NELF association and function.

151 n of NELF-A, -C, or -E, the vast majority of NELF-regulated genes were down-regulated.

152 nism of P-TEFb recruitment and regulation of NELF/DSIF during transcription is not fully understood.

153 ronal enhancers impairs transient release of NELF from the specific target promoters during transcrip

154 The amino acid sequences of NELF-B and NELF-D are highly conserved throughout their

155 To determine the full spectrum of NELF target genes in Drosophila, we performed a microarr

156 Surprisingly, only one-third of NELF target genes were, like Hsp70, up-regulated by NELF

157 or mutation of the ADP-ribosylation sites on NELF-E promotes Pol II pausing, providing a clear functi

158 Depletion of one NELF subunit in salivary glands using RNA interference a

159 ow report new activities of NELF-B and other NELF complex subunits, which are to attenuate glucocorti

160 als that recruitment of COBRA1 and the other NELF subunits to endogenous ERalpha-responsive promoters

161 orchestrates efficient pausing by recruiting NELF to promoters before transcription initiation and by

162 nascent transcript and subsequently recruits NELF.

163 P-TEFb-mediated phosphorylation of Spt5, NELF-A and NELF-E results in the dissociation of NELF fr

164 o binds RNA in vivo, and anchors the subunit NELF-E, which is confirmed to bind RNA in vivo.

165 plex formed by conserved regions in subunits NELF-A and NELF-C, and resolve its crystal structure.

166 Surprisingly, NELF associates with almost one-half of the most highly

167 e LTR showed that pol II was paused and that NELF depletion released pol II.

168 These results demonstrate that NELF plays a role as a common guidance molecule for olfa

169 omatin immunoprecipitation demonstrated that NELF, a negative transcription elongation factor, was as

170 2 cells depleted of NELF and discovered that NELF RNAi affects many rapidly inducible genes involved

171 The finding that NELF also associates with the promoter regions of wingle

172 Lastly, our study indicates that NELF regulates alternative transcription initiation of B

173 most highly expressed genes, indicating that NELF is not necessarily a repressor of gene expression.

174 We propose that NELF and DSIF cause polymerase to pause in the promoter

175 We propose that NELF-mediated pausing allows Pol II to direct CBP-mediat

176 ChIP assays reveal that NELF-B diminishes GR recruitment to promoter regions of

177 ray chip [ChIP-chip] analysis) revealed that NELF is concentrated at the 5' ends of 2,111 genes in Dr

178 n vivo protein-DNA cross-linking showed that NELF and DSIF associate with the promoter region before

179 Chromatin immunoprecipitation showed that NELF knockdown led to dissociation of RNAPII from the pr

180 ted the cross-linking result and showed that NELF, DSIF, and RNA polymerase IIa colocalize at the hsp

181 The NELF-AC subcomplex binds single-stranded nucleic acids i

182 ase pausing, in contrast to depletion of the NELF (negative elongation factor) pausing complex.

183 ction in a manner that is independent of the NELF complex.

184 found to bind to NELF-E, a component of the NELF complex.

185 nctions with TEAD to regulate binding of the NELF negative elongation factor and block SMAD2,3 induct

186 tic relationship between the presence of the NELF pausing factor and positioning of the +1 nucleosome

187 NA binding, whereas the opposite face of the NELF-AC subcomplex binds NELF-B.

188 osomes shows extensive colocalization of the NELF-B and NELF-D subunits at hundreds of interbands.

189 g small hairpin RNA (shRNA) knockdown of the NELF-E subunit, a condition that induced high levels of

190 ions additively affected the affinity of the NELF-E-binding aptamer, whose interaction depended mainl

191 get gene transcription and interact with the NELF complex.

192 BRD4 is also found to bind to NELF-E, a component of the NELF complex.

193 and Tat-SF1, whereas SEC recruits P-TEFb to NELF-A and NELF-E via Paf1c and Med26, respectively.

194 modulators of GR-regulated transactivation, NELF-A and NELF-B, relative to other factors in the over

195 Unexpectedly, NELF-E modifies GR induction in a manner that is indepen

196 enes showed decreased expression levels upon NELF RNAi.

197 use is influenced by the timing between when NELF loads onto Pol II and how fast Pol II escapes the p

198 We propose a model in which NELF recruits Pcf11 and NCoR1-GPS2-HDAC3 to paused RNAP

199 ly conserved throughout their lengths, while NELF-A and NELF-E contain nonconserved regions inserted

200 hat haploinsufficiency of SLBP and/or WHSC2 (NELF-A) contributes to several novel cellular phenotypes

201 determine if paused Pol II colocalized with NELF.

202 Forty-six of 56 genes with NELF were found to have paused Pol II.

203 BRCA1-interacting protein and subunit (with NELF-A, -C/D, and -E) of the human negative elongation f