PubMed ID

18411249

UMMS Affiliation

Department of Cell Biology

Date

4-16-2008

Document Type

Article

Subjects

Adenosine Triphosphate; Antigens, Nuclear; Asparagine; Cytoplasm; DEAD-box RNA Helicases; Fluorescence Recovery After Photobleaching; Hela Cells; Humans; Lysine; Mitosis; Mutant Proteins; Nuclear Matrix-Associated Proteins; Point Mutation; Protein Binding; RNA Splicing; *RNA Transport; RNA, Messenger; RNA-Binding Proteins

Disciplines

Cell Biology | Life Sciences | Medicine and Health Sciences

Abstract

The major-histocompatibility-complex protein UAP56 (BAT1) is a DEAD-box helicase that is deposited on mRNA during splicing. UAP56 is retained on spliced mRNA in an exon junction complex (EJC) or, alternatively, with the TREX complex at the 5' end, where it might facilitate the export of the spliced mRNA to the cytoplasm. Using confocal microscopy, UAP56 was found to be concentrated in RNA-splicing speckled domains of nuclei but was also enriched in adjacent nuclear regions, sites at which most mRNA transcription and splicing occur. At speckled domains, UAP56 was in complexes with the RNA-splicing and -export protein SRm160, and, as measured by FRAP, was in a dynamic binding equilibrium. The application of an in vitro FRAP assay, in which fluorescent nuclear proteins are photobleached in digitonin-extracted cells, revealed that the equilibrium binding of UAP56 in complexes at speckled domains was directly regulated by ATP binding. This was confirmed using a point mutant of UAP56 that did not bind ATP. Point mutation of UAP56 to eliminate ATP binding did not affect RNA splicing, but strongly inhibited the export of mRNA to the cytoplasm.

Rights and Permissions

Citation: J Cell Sci. 2008 May 1;121(Pt 9):1526-37. Epub 2008 Apr 14. Link to article on publisher's site

Related Resources

Link to Article in PubMed