Document Detail


Engineering and characterization of the ribosomal L10.L12 stalk complex. A structural element responsible for high turnover of the elongation factor G-dependent GTPase.
MedLine Citation:
PMID:  18936095     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The ribosomal stalk protein L12 is essential for events dependent on the GTP-binding translation factors. It has been recently shown that ribosomes from Thermus thermophilus contain a heptameric complex L10.(L12)2.(L12)2.(L12)2, rather than the conventional pentameric complex L10.(L12)2.(L12)2. Here we describe the reconstitution of the heptameric complex from purified L10 and L12 and the characterization of its role in elongation factor G-dependent GTPase activity using a hybrid system with Escherichia coli ribosomes. The T. thermophilus heptameric complex resulted in a 2.5-fold higher activity than the E. coli pentameric complex. The structural element of the T. thermophilus complex responsible for the higher activity was investigated using a chimeric L10 protein (Ec-Tt-L10), in which the C-terminal L12-binding site in E. coli L10 was replaced with the same region from T. thermophilus, and two chimeric L12 proteins: Ec-Tt-L12, in which the E. coli N-terminal domain was fused with the T. thermophilus C-terminal domain, and Tt.Ec-L12, in which the T. thermophilus N-terminal domain was fused with the E. coli C-terminal domain. High GTPase turnover was observed with the pentameric chimeric complex formed from E. coli L10 and Ec-Tt-L12 but not with the heptameric complex formed from Ec-Tt-L10 and Tt.Ec-L12. This suggested that the C-terminal region of T. thermophilus L12, rather than the heptameric nature of the complex, was responsible for the high GTPase turnover. Further analyses with other chimeric L12 proteins identified helix alpha6 as the region most likely to contain the responsible element.
Authors:
Tomohiro Miyoshi; Takaomi Nomura; Toshio Uchiumi
Related Documents :
2657655 - The histone-like h protein of escherichia coli is ribosomal protein s3.
18936095 - Engineering and characterization of the ribosomal l10.l12 stalk complex. a structural e...
4929285 - Cotransduction of stra and ribosomal protein cistrons in escherichia coli-salmonella ty...
7007045 - Functional studies on ribosomes lacking protein l1 from mutant escherichia coli.
17342185 - Helping wingless take flight: how wnt proteins are secreted.
12239625 - Somatic mutation and snp in the promoter of dbpa and human hepatocarcinogenesis.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-10-20
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  284     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2008-12-29     Completed Date:  2009-02-24     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  85-92     Citation Subset:  IM    
Affiliation:
Department of Biology, Faculty of Science, Niigata University, Niigata 950-2181, Japan.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics,  metabolism*
Escherichia coli / genetics,  metabolism
Multiprotein Complexes / genetics,  metabolism*
Peptide Elongation Factor G / genetics,  metabolism*
Protein Structure, Quaternary / genetics
Protein Structure, Tertiary / genetics
Recombinant Fusion Proteins / genetics,  metabolism
Ribosomal Proteins / genetics,  metabolism*
Ribosomes / genetics,  metabolism*
Thermus thermophilus / genetics,  metabolism*
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Multiprotein Complexes; 0/Peptide Elongation Factor G; 0/Recombinant Fusion Proteins; 0/Ribosomal Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Identification and role of the homodimerization interface of the glycosylphosphatidylinositol-anchor...
Next Document:  Assembly of high order G alpha q-effector complexes with RGS proteins.