Robust mRNA transcription in chicken DT40 cells depleted of TAFII31 suggests both functional degeneracy and evolutionary divergence

被引:31
作者
Chen, Z [1 ]
Manley, JL [1 ]
机构
[1] Columbia Univ, Dept Biol Sci, New York, NY 10027 USA
关键词
D O I
10.1128/MCB.20.14.5064-5076.2000
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
We have employed gene targeting coupled with conditional expression to construct a chicken DT40 cell line in which a tetracycline (Tet)-repressible promoter is exclusively responsible for expression of cTAF(II)31, a histone-like TAF(II) residing in both the transcription factor TFIID and the histone acetylase complex PCAF/ SAGA. Tet addition resulted in rapid loss of cTAF(II)31 mRNA and protein, eventually leading to apoptotic cell death. Significantly, five of six other TAF(II)s tested were also rapidly depleted, but levels of the TATA binding protein and subunits of PCAF/SAGA were at most modestly compromised. Strikingly, pulse-labeling experiments indicate that total poly(A)(+) mRNA transcription was not significantly reduced after cTAF(II)31 depiction, and steady-state levels of several specific transcripts remained the same or decreased only mildly. Moreover, activation of c-fos transcription following serum starvation occurred efficiently in the absence of cTAF(II)31. These data, which contrast with comparable studies in yeast, strongly suggest that cTAF(II)31 and perhaps other TAF(II)s are not essential for general mRNA transcription in DT10 cells. We propose that this is due to extensive functional degeneracy in the highly complex metazoan transcriptional machinery.
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页码:5064 / 5076
页数:13
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共 70 条
[1]
[Anonymous], 1988, Antibodies: A Laboratory Manual
[2]
Broad, but not universal, transcriptional requirement for yTAFII17, a histone H3-like TAFII present in TFIID and SAGA [J].
Apone, LM ;
Virbasius, CA ;
Holstege, FCP ;
Wang, J ;
Young, RA ;
Green, MR .
MOLECULAR CELL, 1998, 2 (05) :653-661
[3]
Regulation of gene expression by multiple forms of TFIID and other novel TAFII-containing complexes [J].
Bell, B ;
Tora, L .
EXPERIMENTAL CELL RESEARCH, 1999, 246 (01) :11-19
[4]
Human TAFII28 and TAFII18 interact through a histone fold encoded by atypical evolutionary conserved motifs also found in the SPT3 family [J].
Birck, C ;
Poch, O ;
Romier, C ;
Ruff, M ;
Mengus, G ;
Lavigne, AC ;
Davidson, I ;
Moras, D .
CELL, 1998, 94 (02) :239-249
[5]
Mammalian Srb Mediator complex is targeted by adenovirus E1A protein [J].
Boyer, TG ;
Martin, MED ;
Lees, E ;
Ricciardi, RP ;
Berk, AJ .
NATURE, 1999, 399 (6733) :276-279
[6]
The downstream core promoter element, DPE, is conserved from Drosophila to humans and is recognized by TAF(II)60 of Drosophila [J].
Burke, TW ;
Kadonaga, JT .
GENES & DEVELOPMENT, 1997, 11 (22) :3020-3031
[7]
Biochemistry and structural biology of transcription factor IID (TFIID) [J].
Burley, SK ;
Roeder, RG .
ANNUAL REVIEW OF BIOCHEMISTRY, 1996, 65 :769-799
[8]
UNIQUE TATA-BINDING PROTEIN-CONTAINING COMPLEXES AND COFACTORS INVOLVED IN TRANSCRIPTION BY RNA POLYMERASE-II AND POLYMERASE-III [J].
CHIANG, CM ;
GE, H ;
WANG, ZX ;
HOFFMANN, A ;
ROEDER, RG .
EMBO JOURNAL, 1993, 12 (07) :2749-2762
[9]
mRNA capping enzyme is recruited to the transcription complex by phosphorylation of the RNA polymerase II carboxy-terminal domain [J].
Cho, EJ ;
Takagi, T ;
Moore, CR ;
Buratowski, S .
GENES & DEVELOPMENT, 1997, 11 (24) :3319-3326
[10]
SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159