Distribution of B52 within a chromosomal locus depends on the level of transcription.
Drosophila B52 protein is a homologue of human ASF/SF2 that functions in vitro as an essential pre-mRNA splicing factor. Immunofluorescence analysis of polytene chromosomes has shown that B52 generally colocalizes with RNA polymerase II; however, in contrast to other splicing factors, B52 brackets RNA polymerase II at highly active heat-shock puffs. Also, UV cross-linking in nonpolytene cells has shown that B52 cross-links in vivo to DNA flanking the highly active transcription units. Here, we find that the distribution of cross-linked B52 at heat-shock loci depends on transcription levels. Heat shocks at low and moderate temperatures, which induce corresponding levels of transcription, recruit B52 both to transcribed DNA and to flanking DNA, whereas a full heat-shock induction concentrates B52 on the DNA that brackets the entire activated region. We have also identified a 46-kDa protein from Chironomus tentans that binds Drosophila B52 antibodies and has a distribution on chromosomes analogous to B52. This protein is found throughout the moderately transcribed Balbiani rings. However, when transcription at these rings is hyperinduced to levels comparable to fully induced Drosophila heat-shock genes, the protein is restricted to the boundaries of highly decondensed chromatin. We suggest that B52 tracks to chromatin fibers that are folding or unfolding, and we discuss this in light of B52's proposed roles in pre-mRNA splicing and control.
Champlin
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DT
Section of Biochemistry, Cornell University, Ithaca, New York 14853.
Lis
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002683:Chironomidae / Q000276:immunology / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure; D002875:Chromosomes / Q000378:metabolism*; D003429:Cross Reactions; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D007301:Insect Hormones / Q000032:analysis* / Q000276:immunology; D009687:Nuclear Proteins / Q000737:chemistry; D010750:Phosphoproteins; D010862:Pilocarpine / Q000494:pharmacology; D012319:RNA Polymerase II / Q000378:metabolism; D012322:RNA Precursors / Q000378:metabolism; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; D012689:Sequence Homology, Nucleic Acid; D000068103:Serine-Arginine Splicing Factors; D014158:Transcription, Genetic / Q000187:drug effects
D000911:Antibodies, Monoclonal; D002843:Chromatin; D029721:Drosophila Proteins; D006360:Heat-Shock Proteins; D007301:Insect Hormones; D009687:Nuclear Proteins; D010750:Phosphoproteins; D012322:RNA Precursors; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; C489470:SRSF6 protein, human; D010862:Pilocarpine; C070791:B52 protein, Drosophila; D000068103:Serine-Arginine Splicing Factors; D012319:RNA Polymerase II