A protein of the SR family of splicing factors binds extensively to exonic Balbiani ring pre-mRNA and accompanies the RNA from the gene to the nuclear pore.
We report on the molecular cloning and intracellular localization of a heterogeneous nuclear ribonucleoprotein (hnRNP), Ct-hrp45, one of the major components of pre-mRNP particles in Chironomus tentans. It is shown that hrp45 belongs to the SR family of splicing factors and exhibits high sequence similarity to Drosophila SRp55/B52 and human SF2/ASF. The distribution of hrp45 within the C. tentans salivary gland cells is studied by immunocytology. The hrp45 protein is found to be abundant in the nucleus, whereas it is undetectable in the cytoplasm. The fate of hrp45 in specific pre-mRNP particles, the Balbiani ring (BR) granules, is revealed by immunoelectron microscopy. It is observed that hrp45 is associated with the growing BR pre-mRNP particles and is being added continuously concomitant with the growth of the transcript, indicating that hrp45 is bound extensively to exon 4, which comprises 80-90% of the primary transcript. Furthermore, hrp45 remains bound to the BR RNP particles in the nucleoplasm and is not released until the particles translocate through the nuclear pore. Thus, hrp45 behaves as an hnRNP protein linked to exon RNA (and perhaps also to the introns) rather than as a spliceosome component connected to the assembly and disassembly of spliceosomes. It seems that hrp45, and possibly also other SR family proteins, is playing an important role in the structural organization of pre-mRNP particles and is perhaps participating not only in splicing but also in other intranuclear events.
Alzhanova-Ericsson
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AT
Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, Stockholm, Sweden.
Sun
X
X
Visa
N
N
Kiseleva
E
E
Wurtz
T
T
Daneholt
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism*; D002683:Chironomidae; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D019476:Insect Proteins; D016253:Microscopy, Immunoelectron; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009685:Nuclear Envelope; D009687:Nuclear Proteins / Q000235:genetics; D010750:Phosphoproteins; D011485:Protein Binding; D011506:Proteins / Q000235:genetics; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012469:Salivary Glands / Q000737:chemistry / Q000166:cytology; D017386:Sequence Homology, Amino Acid; D000068103:Serine-Arginine Splicing Factors; D014158:Transcription, Genetic
D018076:DNA, Complementary; C102300:HRP45 protein, Chironomus tentans; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D019476:Insect Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D011506:Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C489470:SRSF6 protein, human; D000068103:Serine-Arginine Splicing Factors