Two zinc finger proteins from Xenopus laevis bind the same region of 5S RNA but with different nuclease protection patterns.
Immature oocytes from Xenopus laevis contain a 42S ribonucleoprotein particle (RNP) containing 5S RNA, tRNA, a 43 kDa protein, and a 48 kDa protein. A particle containing 5S RNA and the 43 kDa protein (p43-5S) liberated from the 42S particle upon brief treatment with urea can be purified by anion exchange chromatography. The purified p43-5S RNA migrates as a distinct species during electrophoresis on native polyacrylamide gels. Radiolabeled 5S RNA can be incorporated into the p43-5S complex by an RNA exchange reaction. The resulting complexes containing labeled 5S RNA have a mobility on polyacrylamide gels identical to that of purified p43-5S RNPs. RNP complexes containing 5S RNA labeled at either the 5' or 3' end were probed with a variety of nucleases in order to identify residues protected by p43. Nuclease protection assays performed with alpha-sarcin indicate that p43 binds primarily helices I, II, IV, and V of 5S RNA. This is the same general binding site observed for TFIIIA on 5S RNA. Direct comparison of the binding sites of p43 and TFIIIA with T1 and cobra venom nucleases reveals striking differences in the protection patterns of these two proteins.
Sands
M S
MS
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794.
Bogenhagen
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DF
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002852:Chromatography, Ion Exchange; D004722:Endoribonucleases; D005656:Fungal Proteins; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012341:RNA, Ribosomal, 5S / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000378:metabolism; D014982:Xenopus laevis; D016335:Zinc Fingers
D005656:Fungal Proteins; D012341:RNA, Ribosomal, 5S; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors; C014801:alpha-sarcin; D004722:Endoribonucleases; D012260:Ribonucleases