Characterization of the calf thymus hairpin-binding factor involved in histone pre-mRNA 3' end processing.
Using ion exchange chromatography we have enriched the RNA hairpin-binding factor involved in histone pre-mRNA processing from calf thymus whole cell extract. We demonstrate that the interaction of the factor with its target RNA sequence, the hairpin structure located at the 3' end of mature histone mRNA, is sequence-specific and highly salt-resistant. We have developed a simple in vitro system which allows detection of activities stimulating histone pre-mRNA 3' end processing, based on mouse cell nuclear extract fractionated by Mono Q column chromatography. Using this system, we show that the bovine hairpin-binding factor participates in histone pre-mRNA 3' end processing in vitro. We have further purified the hairpin-binding factor in form of a RNA.protein complex by RNA-mediated elution from phosphocellulose. This led to a fraction highly enriched for 2 proteins of 40 and 43 kDa.
Schaller
A
A
Abteilung Entwicklungsbiologie, Zoologisches Institut, Universität Bern, Baltzerstrasse 4, CH 3012 Bern, Switzerland.
Martin
F
F
Müller
B
B
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002845:Chromatography; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D006657:Histones / Q000096:biosynthesis*; D007700:Kinetics; D051379:Mice; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides; D012322:RNA Precursors / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013950:Thymus Gland / Q000378:metabolism*
D006657:Histones; D009843:Oligoribonucleotides; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D017886:Durapatite