Transcription-dependent redistribution of the large subunit of RNA polymerase II to discrete nuclear domains.
A subpopulation of the largest subunit of RNA polymerase II (Pol II LS) is located in 20-50 discrete subnuclear domains that are closely linked to speckle domains, which store splicing proteins. The speckle-associated fraction of Pol II LS is hyperphosphorylated on the COOH-terminal domain (CTD), and it is highly resistant to extraction by detergents. A diffuse nucleoplasmic fraction of Pol II LS is relatively hypophosphorylated on the CTD, and it is easily extracted by detergents. In transcriptionally active nuclei, speckle bound hyperphosphorylated Pol II LS molecules are distributed in irregularly shaped speckle domains, which appear to be interconnected via a reticular network. When transcription is inhibited, hyperphosphorylated Pol II LS and splicing protein SC35 accumulate in speckle domains, which are transformed into enlarged, dot-like structures lacking interconnections. When cells are released from transcriptional inhibition, Pol IIO and SC35 redistribute back to the interconnected speckle pattern of transcriptionally active cells. The redistribution of Pol II and SC35 is synchronous, reversible, and temperature dependent. It is concluded that: (a) hyperphosphorylation of Pol II LS's CTD is a better indicator of its tight association to discrete subnuclear domains than its transcriptional activity; (b) during states of transcriptional inhibition, hyperphosphorylated Pol II LS can be stored in enlarged speckle domains, which under the light microscope appear to coincide with the storage sites for splicing proteins; and (c) Pol II and splicing proteins redistribute simultaneously according to the overall transcriptional activity of the nucleus.
Bregman
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DB
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Du
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L
van der Zee
S
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Warren
S L
SL
D000546:Amanitins / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology* / Q000378:metabolism; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D000939:Epitopes / Q000032:analysis; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D012319:RNA Polymerase II / Q000032:analysis / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D012261:Ribonucleoproteins; D017381:Spliceosomes / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic / Q000187:drug effects
D000546:Amanitins; D000911:Antibodies, Monoclonal; D000939:Epitopes; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D004004:Dichlororibofuranosylbenzimidazole; D012319:RNA Polymerase II