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Published 5 August 2002. doi:10.1083/jcb.200202035
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© The Rockefeller University Press, 0021-9525/2002/8/463 $5.00
The Journal of Cell Biology, Volume 158, Number 3, August 5, 2002 463-473

Article

Nuclear DNA helicase II is recruited to IFN-{alpha}–activated transcription sites at PML nuclear bodies



Beata Fuchsová1, Petr Novák2, Jarmila Kafková3 and Pavel Hozák1,4

1 Department of Cell Ultrastructure and Molecular Biology, Institute of Experimental Medicine
2 Mass Spectrometry Laboratory, Institute of Microbiology, Academy of Sciences of the Czech Republic, 142 20 Prague 4, Czech Republic
3 Department of Immunology, Insititute of Rheumatology, 128 50 Prague 2, Czech Repulic
4 Department of Cell and Molecular Biology, Third Medical Faculty, Charles University, 100 34 Prague 2, Czech Republic

Address correspondence to Pavel Hozák, Institute of Experimental Medicine, Vídeská 1083, 142 20 Prague 4, Czech Republic. Tel./Fax: 420-2-4016-2219. E-mail: hozak{at}biomed.cas.cz

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.

Key Words: interferon; nuclear DNA helicase II; PML nuclear body; RNA helicase A; RNA polymerase II


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