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Molecular characterization of Drosophila NELF

NELF and DSIF act together to inhibit transcription elongation in vitro, and are implicated in causing promoter proximal pausing on the hsp70 gene in Drosophila. Here, further characterization of Drosophila NELF is provided. Drosophila NELF has four subunits similar to subunits of human NELF. The am...

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Autores principales: Wu, Chwen-Huey, Lee, Chanhyo, Fan, Ruopeng, Smith, Meghan J., Yamaguchi, Yuki, Handa, Hiroshi, Gilmour, David S.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC552961/
https://www.ncbi.nlm.nih.gov/pubmed/15741180
http://dx.doi.org/10.1093/nar/gki274
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author Wu, Chwen-Huey
Lee, Chanhyo
Fan, Ruopeng
Smith, Meghan J.
Yamaguchi, Yuki
Handa, Hiroshi
Gilmour, David S.
author_facet Wu, Chwen-Huey
Lee, Chanhyo
Fan, Ruopeng
Smith, Meghan J.
Yamaguchi, Yuki
Handa, Hiroshi
Gilmour, David S.
author_sort Wu, Chwen-Huey
collection PubMed
description NELF and DSIF act together to inhibit transcription elongation in vitro, and are implicated in causing promoter proximal pausing on the hsp70 gene in Drosophila. Here, further characterization of Drosophila NELF is provided. Drosophila NELF has four subunits similar to subunits of human NELF. The amino acid sequences of NELF-B and NELF-D are highly conserved throughout their lengths, while NELF-A and NELF-E contain nonconserved regions inserted between conserved N- and C-terminal regions. Immunodepletion of NELF or DSIF from a nuclear extract desensitizes transcription in vitro to DRB. Immunodepletion of NELF also impairs promoter proximal pausing on the hsp70 promoter in vitro without affecting initiation. Chromatin immunoprecipitation analyses detect NELF at the promoters of the hsp70 and β1-tubulin genes where promoter proximal pausing has been previously detected. Heat shock induction of hsp70 results in a marked decrease in NELF at the hsp70 promoter. Immunofluorescence analysis of polytene chromosomes shows extensive colocalization of the NELF-B and NELF-D subunits at hundreds of interbands. Neither subunit appears to be recruited to puffs. These results provide a foundation for genetic and biochemical analysis of NELF in Drosophila.
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spelling pubmed-5529612005-03-10 Molecular characterization of Drosophila NELF Wu, Chwen-Huey Lee, Chanhyo Fan, Ruopeng Smith, Meghan J. Yamaguchi, Yuki Handa, Hiroshi Gilmour, David S. Nucleic Acids Res Article NELF and DSIF act together to inhibit transcription elongation in vitro, and are implicated in causing promoter proximal pausing on the hsp70 gene in Drosophila. Here, further characterization of Drosophila NELF is provided. Drosophila NELF has four subunits similar to subunits of human NELF. The amino acid sequences of NELF-B and NELF-D are highly conserved throughout their lengths, while NELF-A and NELF-E contain nonconserved regions inserted between conserved N- and C-terminal regions. Immunodepletion of NELF or DSIF from a nuclear extract desensitizes transcription in vitro to DRB. Immunodepletion of NELF also impairs promoter proximal pausing on the hsp70 promoter in vitro without affecting initiation. Chromatin immunoprecipitation analyses detect NELF at the promoters of the hsp70 and β1-tubulin genes where promoter proximal pausing has been previously detected. Heat shock induction of hsp70 results in a marked decrease in NELF at the hsp70 promoter. Immunofluorescence analysis of polytene chromosomes shows extensive colocalization of the NELF-B and NELF-D subunits at hundreds of interbands. Neither subunit appears to be recruited to puffs. These results provide a foundation for genetic and biochemical analysis of NELF in Drosophila. Oxford University Press 2005 2005-03-01 /pmc/articles/PMC552961/ /pubmed/15741180 http://dx.doi.org/10.1093/nar/gki274 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Article
Wu, Chwen-Huey
Lee, Chanhyo
Fan, Ruopeng
Smith, Meghan J.
Yamaguchi, Yuki
Handa, Hiroshi
Gilmour, David S.
Molecular characterization of Drosophila NELF
title Molecular characterization of Drosophila NELF
title_full Molecular characterization of Drosophila NELF
title_fullStr Molecular characterization of Drosophila NELF
title_full_unstemmed Molecular characterization of Drosophila NELF
title_short Molecular characterization of Drosophila NELF
title_sort molecular characterization of drosophila nelf
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC552961/
https://www.ncbi.nlm.nih.gov/pubmed/15741180
http://dx.doi.org/10.1093/nar/gki274
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