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Molecular and Cellular Biology, April 2004, p. 3324-3336, Vol. 24, No. 8
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.8.3324-3336.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Human Spt6 Stimulates Transcription Elongation by RNA Polymerase II In Vitro

Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8501,¹ The Center for Integrative Bioscience, Okazaki National Research Institutes, Myodaiji, Okazaki 444-8585,² Tokyo Research Laboratories, Kyowa Hakko Kogyo, Machida, Tokyo 194-8533,³ PRESTO, Japan Science and Technology Corporation, Yokohama 226-8503, Japan,⁴ Howard Hughes Medical Institute, Division of Nucleic Acids Enzymology, Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854⁵

Received 10 September 2003/ Returned for modification 16 October 2003/ Accepted 11 December 2003

Recent studies have suggested that Spt6 participates in the regulation of transcription by RNA polymerase II (RNAPII). However, its underlying mechanism remains largely unknown. One possibility, which is supported by genetic and biochemical studies of Saccharomyces cerevisiae, is that Spt6 affects chromatin structure. Alternatively, Spt6 directly controls transcription by binding to the transcription machinery. In this study, we establish that human Spt6 (hSpt6) is a classic transcription elongation factor that enhances the rate of RNAPII elongation. hSpt6 is capable of stimulating transcription elongation both individually and in concert with DRB sensitivity-inducing factor (DSIF), comprising human Spt5 and human Spt4. We also provide evidence showing that hSpt6 interacts with RNAPII and DSIF in human cells. Thus, in vivo, hSpt6 may regulate multiple steps of mRNA synthesis through its interaction with histones, elongating RNAPII, and possibly other components of the transcription machinery.

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