Nuclear DNA helicase II is recruited to IFN-alpha-activated transcription sites at PML nuclear bodies.
It is known that nuclear DNA helicase II (NDH II) links CREB-binding protein directly to RNA polymerase II holoenzyme, and that this interaction is essential for gene activation by CREB. Here, we report for the first time that some NDH II/RNA helicase A is a component of promyelocytic leukemia nuclear bodies (PML NBs). An autoimmune serum specific for PML NBs was identified and used in immunoprecipitation experiments. NDH II was present in the immunoprecipitates as shown by mass spectrometry and by immunoblotting. Immunofluorescence and ultrastructural studies showed that NDH II colocalizes with a small subset of PML NBs in control cells, however, colocalizes with practically all bodies in interferon-alpha-stimulated cells. After interferon stimulation, more PML NBs were found to contain newly synthesized RNA, as indicated by bromouridine incorporation. PML NBs also contain RNA polymerase II. The association of NDH II with PML NBs was transcriptionally dependent, and NDH II was present in all bodies with nascent RNA. Blocking of mRNA synthesis caused NDH II relocalization from nucleoplasm to nucleoli. Based on the data, we suggest that NDH II recruitment to PML NBs is connected with transcriptional regulation of interferon-alpha-inducible genes attached to PML NBs.
Fuchsová
Beata
B
Department of Cell Ultrastructure and Molecular Biology, Institute of Experimental Medicine, Academy of the Czech Republic, Czech Republic.
Novák
Petr
P
Kafková
Jarmila
J
Hozák
Pavel
P
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000546:Amanitins / Q000494:pharmacology; D001327:Autoimmune Diseases / Q000276:immunology; D001798:Blood Proteins / Q000276:immunology / Q000494:pharmacology; D022003:Cell Nucleus Structures / Q000201:enzymology* / Q000648:ultrastructure; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000201:enzymology* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D016898:Interferon-alpha / Q000235:genetics* / Q000494:pharmacology; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D011233:Precipitin Tests; D000071156:Promyelocytic Leukemia Protein; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics*; D025521:Tumor Suppressor Proteins
D000546:Amanitins; D001798:Blood Proteins; D016898:Interferon-alpha; D046911:Macromolecular Substances; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D000071156:Promyelocytic Leukemia Protein; D012333:RNA, Messenger; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D004247:DNA; D000251:Adenosine Triphosphatases; D004265:DNA Helicases