HEL/UAP56 binds cotranscriptionally to the Balbiani ring pre-mRNA in an intron-independent manner and accompanies the BR mRNP to the nuclear pore.
The splicing factor UAP56/HEL/Sub2p is essential for mRNA export. It has been proposed that UAP56/HEL/Sub2p interacts with the pre-mRNA during splicing and recruits the export factor Aly/REF/Yra1 (reviewed in ) to the spliced mRNA. However, UAP56/HEL/Sub2p also participates in the transport of intronless mRNAs, and thus its role in export is not necessarily coupled to splicing. Here, we characterize the HEL protein of Chironomus tentans and we analyze in situ the interaction of HEL with a natural export substrate, the Balbiani ring pre-messenger ribonucleoprotein (BR pre-mRNP, reviewed in ). Using immunoelectron microscopy, we show that HEL binds to the BR pre-mRNP cotranscriptionally and that incorporation of HEL into the pre-mRNP is independent of the location of introns along the BR pre-mRNA. We also show that HEL accompanies the BR mRNP to the nuclear pore and is released from the BR mRNP during translocation to the cytoplasm. Aly/REF is also released from the BR mRNP during translocation but after dissociation of HEL. In summary, we have shown that binding of HEL to the BR pre-mRNA occurs independently of splicing, and we have established the point in the export pathway at which HEL and Aly/REF interact with the mRNP.
Kiesler
Eva
E
Department of Molecular Biology and Functional Genomics, University of Stockholm, SE-10691, Stockholm, Sweden.
Miralles
Francesc
F
Visa
Neus
N
D000818:Animals; D001692:Biological Transport; D002683:Chironomidae / Q000235:genetics; D007438:Introns / Q000235:genetics; D016253:Microscopy, Immunoelectron; D022022:Nuclear Pore / Q000378:metabolism* / Q000648:ultrastructure; D011485:Protein Binding; D020365:RNA Helicases / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D012469:Salivary Glands / Q000378:metabolism / Q000648:ultrastructure; D014158:Transcription, Genetic
D012322:RNA Precursors; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D020365:RNA Helicases