Tracking FACT and the RNA polymerase II elongation complex through chromatin in vivo.
RNA polymerase II (Pol II) transcription through nucleosomes is facilitated in vitro by the protein complex FACT (Facilitates Chromatin Transcription). Here we show that FACT is associated with actively transcribed Pol II genes on Drosophila polytene chromosomes. FACT displays kinetics of recruitment and of chromosome tracking in vivo similar to Pol II and elongation factors Spt5 and Spt6. Interestingly, FACT does not colocalize with Pol III-transcribed genes, which are known to undergo nucleosome transfer rather than disassembly in vitro. Our observations are consistent with FACT being restricted to transcription that involves nucleosome disassembly mechanisms.
Saunders
Abbie
A
Molecular Biology and Genetics, Biotechnology Building, Cornell University, Ithaca, NY 14850, USA.
Werner
Janis
J
Andrulis
Erik D
ED
Nakayama
Takahiro
T
Hirose
Susumu
S
Reinberg
Danny
D
Lis
John T
JT
D000818:Animals; D002466:Cell Nucleolus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins / Q000235:genetics; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D000076249:Heat Shock Transcription Factors; D018869:Heat-Shock Response; D006609:High Mobility Group Proteins; D006358:Hot Temperature; D009707:Nucleosomes / Q000378:metabolism; D016366:Open Reading Frames; D010445:Peptide Elongation Factors / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors / Q000378:metabolism*
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018840:HSP70 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D006609:High Mobility Group Proteins; C053134:Hsf protein, Drosophila; D009707:Nucleosomes; D010445:Peptide Elongation Factors; C489543:SSRP protein, Drosophila; C470423:Spt6 protein, Drosophila; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C069940:SPT5 transcriptional elongation factor; D012319:RNA Polymerase II