Identification of a 100-kDa protein associated with nuclear ribonuclease P activity in Schizosaccharomyces pombe.
Ribonuclease P from the fission yeast Schizosaccharomyces pombe has been purified to apparent homogeneity. A purification of 23,000-fold was achieved by four fractionation steps with DEAE-cellulose chromatography, phosphocellulose chromatography, glycerol-gradient fractionation and finally tRNA-affinity chromatography. A 100-kDa protein was present in the most pure preparations in amounts approximately stoichiometric with the previously identified RNA components of the enzyme, K1-RNA and K2-RNA [Krupp, G., Cherayil, B., Frendeway, D., Nishikawa, S. & Söll, D. (1986) EMBO J. 5, 1697-1703]. A cross-linking experiment utilizing a 4-thiouridine-substituted precursor tRNA demonstrated that the 100-kDa protein interacts with the ribonuclease P substrate in a specific fashion. We therefore conclude that the protein component of S. pombe ribonuclease P is a 100-kDa protein.
Zimmerly
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Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114.
Drainas
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Sylvers
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Söll
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D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D004722:Endoribonucleases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D005656:Fungal Proteins / Q000032:analysis*; D008970:Molecular Weight; D012322:RNA Precursors / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012331:RNA, Fungal / Q000032:analysis; D012343:RNA, Transfer / Q000378:metabolism; D011817:Rabbits; D043262:Ribonuclease P; D012568:Schizosaccharomyces / Q000201:enzymology*; D013379:Substrate Specificity
D005656:Fungal Proteins; D012322:RNA Precursors; D016337:RNA, Catalytic; D012331:RNA, Fungal; D012343:RNA, Transfer; D004722:Endoribonucleases; D043262:Ribonuclease P