The elongation factors Pandora/Spt6 and Foggy/Spt5 promote transcription in the zebrafish embryo.
Precise temporal and spatial control of transcription is a fundamental component of embryonic development. Regulation of transcription elongation can act as a rate-limiting step during mRNA synthesis. The mechanisms of stimulation and repression of transcription elongation during development are not yet understood. We have identified a class of zebrafish mutations (pandora, sk8 and s30) that cause multiple developmental defects, including discrete problems with pigmentation, tail outgrowth, ear formation and cardiac differentiation. We demonstrate that the pandora gene encodes a protein similar to Spt6, a proposed transcription elongation factor. Additionally, the sk8 and s30 mutations are null alleles of the foggy/spt5 locus, which encodes another transcription elongation factor. Through real-time RT-PCR analysis, we demonstrate that Spt6 and Spt5 are both required for efficient kinetics of hsp70 transcription in vivo. Altogether, our results suggest that Spt6 and Spt5 play essential roles of comparable importance for promoting transcription during embryogenesis. This study provides the first genetic evidence for parallel functions of Spt6 and Spt5 in metazoans and establishes a system for the future analysis of transcription elongation during development.
Keegan
Brian R
BR
Developmental Genetics Program and Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA.
Feldman
Jessica L
JL
Lee
Diana H
DH
Koos
David S
DS
Ho
Robert K
RK
Stainier
Didier Y R
DY
Yelon
Deborah
D
D000818:Animals; D002874:Chromosome Mapping; D004268:DNA-Binding Proteins; D005260:Female; D018507:Gene Expression Regulation, Developmental; D056488:Histone Chaperones; D006358:Hot Temperature; D008297:Male; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D010445:Peptide Elongation Factors / Q000235:genetics*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D029961:Zebrafish Proteins
D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D009687:Nuclear Proteins; D010445:Peptide Elongation Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C065456:SPT6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D029961:Zebrafish Proteins; C416949:supt5h protein, zebrafish