Accurate processing of human pre-rRNA in vitro

Hannon, G. J., Maroney, P. A., Branch, A., Benenfield, B. J., Robertson, H. D., Nilsen, T. W. (October 1989) Accurate processing of human pre-rRNA in vitro. Molecular and Cellular Biology, 9 (10). pp. 4422-4431. ISSN 02707306 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/2586517
DOI: 10.1128/MCB.9.10.4422

Abstract

We report here that the mature 5' terminus of human 18S rRNA is generated in vitro by a two-step processing reaction. In the first step, SP6 transcripts were specifically cleaved in HeLa cell nucleolar extract at three positions near the external transcribed spacer (ETS)-18S boundary. Of these cleavage sites, two were major and the other was minor. RNase T1 fingerprint and secondary nuclease analyses placed the two major cleavage sites 3 and 8 bases upstream from the mature 5' end of 18S rRNA and the minor cleavage site 1 base into the 18S sequence. All three cleavages yielded 5'-hydroxyl, 2'-3'-cyclic phosphate termini and were 5' of adenosine residues in the sequence UACCU, which was repeated three times near the ETS-18S boundary. In the second step, the initial cleavage product containing 3 bases of ETS was converted to an RNA with a 5' terminus identical to that of mature 18S RNA by an activity found in HeLa cell cytoplasmic extracts.

Item Type: Paper
Uncontrolled Keywords: ribosome rna cell culture hela cell human human cell rna processing Base Sequence Cell Nucleolus Cytoplasm Exoribonucleases Hela Cells Molecular Sequence Data Nucleotide Mapping RNA Precursors RNA Processing, Post-Transcriptional RNA, Ribosomal RNA, Ribosomal, 18S Support, U.S. Gov't, Non-P.H.S. Support, U.S. Gov't, P.H.S.
Subjects: bioinformatics > genomics and proteomics > analysis and processing > DNA RNA Processing
organism description > animal > mammal > primates > hominids > human
CSHL Authors:
Depositing User: Gail Sherman
Date: October 1989
Date Deposited: 27 Jul 2017 18:56
Last Modified: 27 Jul 2017 18:56
PMCID: PMC362525
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34858

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