Evidence for a polynuclear metal ion binding site in the catalytic domain of ribonuclease P RNA

被引：43

作者：

Christian, EL ^{[1
]}

Kaye, NM ^{[1
]}

Harris, ME ^{[1
]}

机构：

[1] Case Western Reserve Univ, Sch Med, Ctr RNA Mol Biol, Cleveland, OH 44106 USA

来源：

EMBO JOURNAL | 2002年 / 21卷 / 09期

关键词：

metal ion interactions; phosphorothioate; ribonuclease P; ribozyme;

D O I：

10.1093/emboj/21.9.2253

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Interactions with divalent metal ions are essential for the folding and function of the catalytic RNA component of the tRNA processing enzyme ribonuclease P (RNase P RNA). However, the number and location of specific metal ion interactions in this large, highly structured RNA are poorly understood. Using atomic mutagenesis and quantitative analysis of thiophilic metal ion rescue we provide evidence for metal ion interactions at the pro-R-P and pro-S-P non-bridging phosphate oxygens at nucleotide A67 in the universally conserved helix P4. Moreover, second-site modifications within helix P4 and the adjacent single stranded region (J3/4) provide the first evidence for metal ion interactions with nucleotide base functional groups in RNase P RNA and reveal the presence of an additional metal ion important for catalytic function. Together, these data are consistent with a cluster of metal ion interactions in the P1-P4 multi-helix junction that defines the catalytic core of the RNase P ribozyme.

引用

页码：2253 / 2262

页数：10

共 49 条

[11] The track of the pre-tRNA 5′ leader in the ribonuclease P ribozyme-substrate complex
Christian, EL
Harris, ME
[J]. BIOCHEMISTRY, 1999, 38 (39) : 12629 - 12638
[12] Helix P4 is a divalent metal ion binding site in the conserved core of the ribonuclease P ribozyme
Christian, EL
Kaye, NM
Harris, ME
[J]. RNA, 2000, 6 (04) : 511 - 519
[13] Metals, motifs, and recognition in the crystal structure of a 5S rRNA domain
Correll, CC
Freeborn, B
Moore, PB
Steitz, TA
[J]. CELL, 1997, 91 (05) : 705 - 712
[14] NUCLEOSIDE PHOSPHOROTHIOATES
ECKSTEIN, F
[J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1985, 54 : 367 - 402
[15] The crystal structure of the dimerization initiation site of genomic HIV-1 RNA reveals an extended duplex with two adenine bulges
Ennifar, E
Yusupov, M
Walter, P
Marquet, R
Ehresmann, B
Ehresmann, C
Dumas, P
[J]. STRUCTURE, 1999, 7 (11) : 1439 - 1449
[16] Feig A. L., 1999, RNA WORLD, P287
[17] Crystal structure of a hepatitis delta virus ribozyme
Ferré-D'Amaré, AR
Zhou, KH
Doudna, JA
[J]. NATURE, 1998, 395 (6702) : 567 - 574
[18] RATIONAL DESIGN OF SELF-CLEAVING PRE-TRANSFER-RNA RIBONUCLEASE-P RNA CONJUGATES
FRANK, DN
HARRIS, ME
PACE, NR
[J]. BIOCHEMISTRY, 1994, 33 (35) : 10800 - 10808
[19] In vitro selection for altered divalent metal specificity in the RNase P RNA
Frank, DN
Pace, NR
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (26) : 14355 - 14360
[20] Kinetic characterization of the second step of group II intron splicing:: Role of metal ions and the cleavage site 2′-OH in catalysis
Gordon, PM
Sontheimer, EJ
Piccirilli, JA
[J]. BIOCHEMISTRY, 2000, 39 (42) : 12939 - 12952

← 1 2 3 4 5 →