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The elongation factors Pandora/Spt6 and Foggy/Spt5 promote transcription in the zebrafish embryo

Brian R. Keegan^1,*, Jessica L. Feldman^1,*, Diana H. Lee¹, David S. Koos², Robert K. Ho², Didier Y. R. Stainier³ and Deborah Yelon^1,

¹ Developmental Genetics Program and Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA
² Department of Molecular Biology, Princeton University, Princeton, NJ, USA
³ Department of Biochemistry and Biophysics and Programs in Developmental Biology, Genetics, and Human Genetics, University of California, San Francisco, San Francisco, CA, USA
^* These authors contributed equally to the work

Author for correspondence (e-mail: yelon{at}saturn.med.nyu.edu)

Accepted 9 January 2002

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.

Supplemental figure available on-line

Key words: Heart, Spt6, Spt5, Transcription elongation, Real-time RT-PCR, hsp70, Zebrafish

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