High-resolution localization of <i>Drosophila</i> Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation

Andrulis, Erik D.; Guzmán, Ernesto; Döring, Pascal; Werner, Janis; Lis, John T.

doi:10.1101/gad.844200

Cited by 248 publications

(251 citation statements)

References 36 publications

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“…10 Mutations in Spt6 influence mRNA 3′-end formation, 11 splicing, and mRNA export. 12 Spt6 is associated with the Pol II machinery, 6,7 , and its SH2 domain is apparently important for this. Spt6 binds to Spt5 4,5 , and occupies the 5′-region of the uninduced heat shock gene hsp70.…”

Section: Introductionmentioning

confidence: 99%

“…Spt6 binds to Spt5 4,5 , and occupies the 5′-region of the uninduced heat shock gene hsp70. 6 In vivo, Spt6 and Spt5 co-localize with actively transcribing Pol II that is phosphorylated at Ser2 of the C-terminal repeat domain (CTD) of the Pol II largest subunit. 7 In vitro, the C-terminal region of mouse Spt6, which includes the SH2 domain, binds the Ser2-phosphorylated CTD, and this interaction is required for normal mRNA processing and exit from the nucleus.…”

Section: Introductionmentioning

confidence: 99%

“…2,3 Like Spt4 and Spt5, Spt6 is required for transcription elongation, 4,5,31,70 and co-localizes with Spt5 and RNA polymerase (Pol) II at transcriptionally active loci on Drosophila polytene chromosomes. 6,7 Spt6 interacts with the structured part of histone H3 and can deposit nucleosomes on DNA in a supercoiling assay. 8 Loss of normal Spt6 function leads to transcription from cryptic promoters within the coding region of genes, 9 suggesting that Spt6 is required to reestablish correct chromatin structure after Pol II passage.…”

Section: Introductionmentioning

confidence: 99%

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Structure and in Vivo Requirement of the Yeast Spt6 SH2 Domain

Dengl

Mayer

Sun

et al. 2009

Journal of Molecular Biology

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During transcription elongation through chromatin, the Ser2-phosphorylated C-terminal repeat domain of RNA polymerase II binds the C-terminal Src homology 2 (SH2) domain of the nucleosome re-assembly factor Spt6. This SH2 domain is unusual in its specificity to bind phosphoserine, rather than phosphotyrosine and because it is the only SH2 domain in the yeast genome. Here, we report the high-resolution crystal structure of the SH2 domain from Candida glabrata Spt6. The structure combines features from both structural subfamilies of SH2 domains, suggesting it resembles a common ancestor of all SH2 domains. Two conserved surface pockets deviate from those of canonical SH2 domains, and may explain the unusual phosphoserine specificity. Differential gene expression analysis reveals that the SH2 domain is required for normal expression of a subset of yeast genes, and is consistent with multiple functions of Spt6 in chromatin transcription.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structure and in Vivo Requirement of the Yeast Spt6 SH2 Domain

Dengl

Mayer

Sun

et al. 2009

Journal of Molecular Biology

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“…In addition, the association of some elongation factors, e.g. FACT, Spt5, Spt6, p-TEFb, etc., are necessary for RNA pol II escaping the paused state and elongating through the entire hsp70 gene [12][13][14]. Many studies have demonstrated that treatments of HDIs can result in histone hyperacetylation and remodeling of chromatin structure.…”

Section: Introductionmentioning

confidence: 99%

Effects of histone deacetylase inhibitors on transcriptional regulation of the hsp70 gene in Drosophila

et al. 2006

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npg Histone acetyltransferases/deacetylases contribute to the activation or inactivation of transcription by modifying the structure of chromatin. Here we examined the effects of histone deacetylase inhibitors (HDIs), trichostatin A, and sodium butyrate on hsp70 gene transcriptional regulation in Drosophila. The chromatin immunoprecipitation assays revealed that HDI treatments induced the hyperacetylation of histone H3 at the promoter and the transcribing regions of hsp70 gene, increased the accessibility of heat-shock factor to target heat-shock element, and promoted the RNA polymerase II-mediated transcription. Moreover, the quantitative real-time PCR confirmed that the HDI-induced hyperacetylation of histone H3 enhanced both the basal and the inducible expression of hsp70 mRNA level. In addition, the acetylation level of histone H3 at the promoter exhibited a fluctuated change upon the time of heat shock. These experimental data implicated a causal link between histone acetylation and enhanced transcription initiation of hsp70 gene in Drosophila.

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“…There is ample genetic and biochemical evidence from budding yeast, zebrafish, roundworm, fruitfly, and mammalian systems that Spt5 and Spt4 exert both negative and positive effects on transcription elongation (Hartzog et al 1998;Andrulis et al 2000;Guo et al 2000;Renner et al 2001;Shim et al 2002;Wu et al 2003;Jennings et al 2004). Spt5 is a large polypeptide (z1000-1200 amino acids) composed of multiple domain modules, including an acidic N-terminal domain, central NusG-like NGN (NusG N-terminal homology), and KOW domains (Hartzog et al 1998;Wade et al 1998a;Ponting 2002), and a CTD targeted for threonine phosphorylation Yamada et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Schizosaccharomyces pombe Spt5-Spt4 complex

Schwer

Schneider²,

Pei³

et al. 2009

RNA

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The Spt5-Spt4 complex regulates early transcription elongation by RNA polymerase II and has an imputed role in pre-mRNA processing via its physical association with mRNA capping enzymes. Here we characterize the Schizosaccharomyces pombe core Spt5-Spt4 complex as a heterodimer and map a trypsin-resistant Spt4-binding domain within the Spt5 subunit. A genetic analysis of Spt4 in S. pombe revealed it to be inessential for growth at 25°C-30°C but critical at 37°C. These results echo the conditional spt4D growth phenotype in budding yeast, where we find that Saccharomyces cerevisiae and S. pombe Spt4 are functionally interchangeable. Complementation of S. cerevisiae spt4D and a two-hybrid assay for Spt4-Spt5 interaction provided a readout of the effects of 33 missense and truncation mutations on S. pombe Spt4 function in vivo, which were interpreted in light of the recent crystal structure of S. cerevisiae Spt4 fused to a fragment of Spt5. Our results highlight the importance of the Spt4 Zn 2+ -binding residues-Cys12, Cys15, Cys29, and Asp32-and of Ser57, a conserved constituent of the Spt4-Spt5 interface. The 990-amino acid S. pombe Spt5 protein has an exceptionally regular carboxyl-terminal domain (CTD) composed of 18 nonapeptide repeats. We find that as few as three nonamer repeats sufficed for S. pombe growth, but only when Spt4 was present. Synthetic lethality of the spt5 1-835 spt4D double mutant at 34°C suggests that interaction of Spt4 with the central domain of Spt5 overlaps functionally with the Spt5 CTD.

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High-resolution localization of Drosophila Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation

Cited by 248 publications

References 36 publications

Structure and in Vivo Requirement of the Yeast Spt6 SH2 Domain

Structure and in Vivo Requirement of the Yeast Spt6 SH2 Domain

Effects of histone deacetylase inhibitors on transcriptional regulation of the hsp70 gene in Drosophila

Characterization of the Schizosaccharomyces pombe Spt5-Spt4 complex

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