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Titolo:
Arrangement of the central pseudoknot region of 16S rRNA in the 30S ribosomal subunit determined by site-directed 4-thiouridine crosslinking
Autore:
Juzumiene, DI; Wollenzien, P;
Indirizzi:
N Carolina State Univ, Dept Biochem, Raleigh, NC 27695 USA N Carolina State Univ Raleigh NC USA 27695 Biochem, Raleigh, NC 27695 USA
Titolo Testata:
RNA-A PUBLICATION OF THE RNA SOCIETY
fascicolo: 1, volume: 7, anno: 2001,
pagine: 71 - 84
SICI:
1355-8382(200101)7:1<71:AOTCPR>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
ESCHERICHIA-COLI; TRANSFER-RNA; CONFORMATIONAL SWITCH; ANGSTROM RESOLUTION; PROTEIN-SYNTHESIS; DECODING SITE; 915 REGION; BINDING; MUTATIONS; NUCLEOTIDES;
Keywords:
16S rRNA three-dimensional structure; intramolecular RNA-RNA crosslinks; mRNA decoding region;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
43
Recensione:
Indirizzi per estratti:
Indirizzo: Wollenzien, P N Carolina State Univ, Dept Biochem, Box 7622, Raleigh, NC 27695 USA N Carolina State Univ Box 7622 Raleigh NC USA 27695 7695 USA
Citazione:
D.I. Juzumiene e P. Wollenzien, "Arrangement of the central pseudoknot region of 16S rRNA in the 30S ribosomal subunit determined by site-directed 4-thiouridine crosslinking", RNA, 7(1), 2001, pp. 71-84

Abstract

The 16S rRNA central pseudoknot region in the 30S ribosomal subunit has been investigated by photocrosslinking from 4-thiouridine (s(4)U) located in the first 20 nt of the 16S rRNA. RNA fragments (nt 1-20) were made by in vitro transcription to incorporate s4U at every uridine position or were madeby chemical synthesis to incorporate s4U into one of the uridine positionsat +5, +14, +17, or +20. These were ligated to RNA containing nt 21-1542 of the 16S rRNA sequence and, after gel purification, the ligated RNA was reconstituted into 30S subunits. Long-range intramolecular crosslinks were produced by near-UV irradiation; these were separated by gel electrophoresis and analyzed by reverse transcription reactions. A number of crosslinks aremade in each of the constructs, which must reflect the structural flexibility or conformational heterogeneity in this part of the 30S subunit. All ofthe constructs show crosslinking to the 559-562, 570-571, and 1080-1082 regions; however, other sites are crosslinked specifically from each s(4)U position. The most distinctive crosslinking sites are: 341-343 and 911-917 for s(4)U(+5); 903-904 (very strong), 1390-1397, and 1492 for s(4)U(+14); and903-904 (moderate) for s4U(+17); in the 1070-1170 region in which there are different patterns for each s(4)U position. These results indicate that part of the central pseudoknot is in close contact with the decoding region,with helix 27 in the 885-912 interval and with part of domain iii RNA. Crosslinking between s(4)U(+14) and 1395-1397 is consistent with base pairing at U14-A1398.

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Documento generato il 27/09/20 alle ore 04:52:09