Efficiency of T4 Gene 60 Translational Bypassing

Please use this identifier to cite or link to this item: http://hdl.handle.net/10045/3377
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Title: Efficiency of T4 Gene 60 Translational Bypassing
Authors: Maldonado, Rafael | Herr, Alan J.
Research Group/s: Transducción de Señales en Bacterias
Center, Department or Service: Universidad de Alicante. Departamento de Fisiología, Genética y Microbiología | University of Utah. Department of Human Genetics
Keywords: Phage | Translation
Knowledge Area: Genética | Microbiología
Issue Date: Apr-1998
Publisher: American Society for Microbiology
Citation: MALDONADO CARO, Rafael Diego; HERR, Alan J. “Efficiency of T4 Gene 60 Translational Bypassing”. Journal of Bacteriology. Vol. 180, No. 7 (Apr. 1998). ISSN 0021-9193, pp. 1822-1830
Abstract: Ribosomes translating bacteriophage T4 gene 60 mRNA bypass 50 noncoding nucleotides from a takeoff site at codon 46 to a landing site just upstream of codon 47. A key signal for efficient bypassing is contained within the nascent peptide synthesized prior to takeoff. Here we show that this signal is insensitive to the addition of coding information at its N terminus. In addition, analysis of amino-terminal fusions, which allow detection of all major products synthesized from the gene 60 mRNA, show that 50% of ribosomes bypass the coding gap while the rest either terminate at a UAG stop codon immediately following codon 46 or fail to resume coding. Bypassing efficiency estimates significantly lower than 50% were obtained with enzymatic reporter systems that relied on comparing test constructs to constructs with a precise excision of the gap (gap deletion). Further analysis showed that these estimates are distorted by differences between test and gap deletion functional mRNA levels. An internal translation initiation site at Met12 of gene 60 (which eliminates part of the essential nascent peptide) also distorts these estimates. Together, these results support an efficiency estimate of ~50%, less than previously reported. This estimate suggests that bypassing efficiency is determined by the competition between reading signals and release factors and gives new insight into the kinetics of bypassing signal action.
Sponsor: This work was supported by the Howard Hughes Medical Institute (via Ray Gesteland) and a grant (to John Atkins) from the NIH (RO1-GM48152-05).
URI: http://hdl.handle.net/10045/3377
ISSN: 0021-9193
Language: eng
Type: info:eu-repo/semantics/article
Peer Review: si
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