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

Citation
Ed. Andrulis et al., High-resolution localization of Drosophila Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation, GENE DEV, 14(20), 2000, pp. 2635-2649
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Cell & Developmental Biology
Journal title
GENES & DEVELOPMENT
ISSN journal
0890-9369 → ACNP
Volume
14
Issue
20
Year of publication
2000
Pages
2635 - 2649
Database
ISI
SICI code
0890-9369(20001015)14:20<2635:HLODSA>2.0.ZU;2-E
Abstract
Recent studies have demonstrated roles for Spt4, Spt5, and Spt6 in the regu lation of transcriptional elongation in both yeast and humans. Here, we sho w that Drosophila Spt5 and Spt6 colocalize at a large number of transcripti onally active chromosomal sites on polytene chromosomes and are rapidly rec ruited to endogenous and transgenic heat shock loci upon heat shock. Costai ning with antibodies to Spt6 and to either the largest subunit of RNA polym erase II or cyclin T, a subunit of the elongation factor P-TEFb, reveals th at all three factors have a similar distribution at sites of active transcr iption. Crosslinking and immunoprecipitation experiments show that Spt5 is present at uninduced heat shock gene promoters, and that upon heat shock, S pt5 and Spt6 associate with the 5' and 3' ends of heat shock genes. Spt6 is recruited within 2 minutes of a heat shock, similar to heat shock factor ( HSF); moreover, this recruitment is dependent on HSF. These findings provid e support for the roles of Spt5 in promoter-associated pausing and of Spt5 and Spt6 in transcriptional elongation in vivo.