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The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a.
J Mol Biol. 2002 Feb 22; 316(3):475-88.JM

Abstract

YL37a is an essential yeast ribosomal protein that has a C(2)-C(2) zinc finger motif. Replacement of the cysteine residues had yielded variants that lacked the capacity to bind zinc but still supported cell growth. In a continuation of an examination of the relation of the structure of YL37a to its function, the contribution of amino acid residues in the intervening sequence between the internal cysteine residues of the motif was evaluated. Substitutions of alanine for the lysine residues at positions 44, 45, or 48, or for arginine 49 slowed cell growth. The most severe effect was caused by a double-mutation, K48A-R49A. A mutation of tryptophan 55 to alanine was lethal. Mutations to alanine of six conserved residues (K6, K7, K13, Y14, R17, and Y18) in the amino-terminal region decreased cell growth; the Y14 mutation was lethal. An in vitro assay for binding of YL37a to individual 26 S rRNA domains was developed. Binding of the recombinant fusion protein MBP-YL37a was to domains II and III; the K(d) for binding to domain II was 79 nM; for domain III it was 198 nM. There was a close correspondence between the effect of mutations in YL37a on cell growth and on binding to 26 S rRNA. In the atomic structure of the 50 S subunit of Haloarcula marismortui, the archaebacteria homolog of yeast YL37a, L37ae, coordinates a zinc atom and the finger motif is folded and interacts mainly with domain III of 23 S rRNA; whereas the amino-terminal region of L37ae interacts primarily with domain II. The biochemical and genetic experiments complement the three-dimensional structure and define for the first time the functional importance of a subset of the residues in close proximity to nucleotides.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11866512

Citation

Dresios, John, et al. "The Role of the Zinc Finger Motif and of the Residues at the Amino Terminus in the Function of Yeast Ribosomal Protein YL37a." Journal of Molecular Biology, vol. 316, no. 3, 2002, pp. 475-88.
Dresios J, Chan YL, Wool IG. The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a. J Mol Biol. 2002;316(3):475-88.
Dresios, J., Chan, Y. L., & Wool, I. G. (2002). The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a. Journal of Molecular Biology, 316(3), 475-88.
Dresios J, Chan YL, Wool IG. The Role of the Zinc Finger Motif and of the Residues at the Amino Terminus in the Function of Yeast Ribosomal Protein YL37a. J Mol Biol. 2002 Feb 22;316(3):475-88. PubMed PMID: 11866512.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a. AU - Dresios,John, AU - Chan,Yuen-Ling, AU - Wool,Ira G, PY - 2002/2/28/pubmed PY - 2002/4/6/medline PY - 2002/2/28/entrez SP - 475 EP - 88 JF - Journal of molecular biology JO - J Mol Biol VL - 316 IS - 3 N2 - YL37a is an essential yeast ribosomal protein that has a C(2)-C(2) zinc finger motif. Replacement of the cysteine residues had yielded variants that lacked the capacity to bind zinc but still supported cell growth. In a continuation of an examination of the relation of the structure of YL37a to its function, the contribution of amino acid residues in the intervening sequence between the internal cysteine residues of the motif was evaluated. Substitutions of alanine for the lysine residues at positions 44, 45, or 48, or for arginine 49 slowed cell growth. The most severe effect was caused by a double-mutation, K48A-R49A. A mutation of tryptophan 55 to alanine was lethal. Mutations to alanine of six conserved residues (K6, K7, K13, Y14, R17, and Y18) in the amino-terminal region decreased cell growth; the Y14 mutation was lethal. An in vitro assay for binding of YL37a to individual 26 S rRNA domains was developed. Binding of the recombinant fusion protein MBP-YL37a was to domains II and III; the K(d) for binding to domain II was 79 nM; for domain III it was 198 nM. There was a close correspondence between the effect of mutations in YL37a on cell growth and on binding to 26 S rRNA. In the atomic structure of the 50 S subunit of Haloarcula marismortui, the archaebacteria homolog of yeast YL37a, L37ae, coordinates a zinc atom and the finger motif is folded and interacts mainly with domain III of 23 S rRNA; whereas the amino-terminal region of L37ae interacts primarily with domain II. The biochemical and genetic experiments complement the three-dimensional structure and define for the first time the functional importance of a subset of the residues in close proximity to nucleotides. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/11866512/The_role_of_the_zinc_finger_motif_and_of_the_residues_at_the_amino_terminus_in_the_function_of_yeast_ribosomal_protein_YL37a_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022283601953744 DB - PRIME DP - Unbound Medicine ER -