Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.

We analyzed protein expression from a cloned Mycoplasma hyorhinis genomic fragment that produces in Escherichia coli a set of related polypeptides of 110, 100, 65, and 55 kilodaltons from a coding region of just over 3.0 kilobases. Expression of these multiple products resulted from a mechanism oper...

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Những tác giả chính: Notarnicola, S M, McIntosh, M A, Wise, K S
Định dạng: Bài viết
Ngôn ngữ:en
Được phát hành: 1990
Những chủ đề:
Research Article
Truy cập trực tuyến:https://ncbi.nlm.nih.gov/pmc/articles/PMC209098/
https://ncbi.nlm.nih.gov/pubmed/2188946
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spelling pubmed-2090982003-12-01 Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli. Notarnicola, S M McIntosh, M A Wise, K S J Bacteriol Research Article We analyzed protein expression from a cloned Mycoplasma hyorhinis genomic fragment that produces in Escherichia coli a set of related polypeptides of 110, 100, 65, and 55 kilodaltons from a coding region of just over 3.0 kilobases. Expression of these multiple products resulted from a mechanism operating at the translational level but not from truncation at UGA termination codons, which are known to encode tryptophan in several mycoplasma species. The structural relatedness of the proteins was demonstrated by two-dimensional tryptic peptic mapping, but their generation by posttranslational processing was ruled out by pulse-chase labeling analysis. Examination of proteins expressed from plasmid constructs and tryptic peptide analysis of these polypeptides and the original set of proteins revealed that they share carboxy-terminal regions, an observation inconsistent with truncation at UGA codons. Expression of proteins from this cloned fragment was not dependent on vector sequences and was observed when the coding region was placed under control of a T7 promoter, suggesting that all products were translated from a single message. Expression of related products in mycoplasmas was examined by immunoblot analysis of M. hyorhinis proteins with antiserum against overexpressed recombinant proteins. A single 115-kilodalton mycoplasma protein was detected, which is larger than any of the related proteins expressed in E. coli. Our analysis indicated that translation initiation sites are used in E. coli that are not active in mycoplasmas, thereby defining differences between the translational regulatory signals of mycoplasmas and eubacteria. 1990-06 /pmc/articles/PMC209098/ /pubmed/2188946 Text en
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Research Article
spellingShingle Research Article
Notarnicola, S M
McIntosh, M A
Wise, K S
Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
description We analyzed protein expression from a cloned Mycoplasma hyorhinis genomic fragment that produces in Escherichia coli a set of related polypeptides of 110, 100, 65, and 55 kilodaltons from a coding region of just over 3.0 kilobases. Expression of these multiple products resulted from a mechanism operating at the translational level but not from truncation at UGA termination codons, which are known to encode tryptophan in several mycoplasma species. The structural relatedness of the proteins was demonstrated by two-dimensional tryptic peptic mapping, but their generation by posttranslational processing was ruled out by pulse-chase labeling analysis. Examination of proteins expressed from plasmid constructs and tryptic peptide analysis of these polypeptides and the original set of proteins revealed that they share carboxy-terminal regions, an observation inconsistent with truncation at UGA codons. Expression of proteins from this cloned fragment was not dependent on vector sequences and was observed when the coding region was placed under control of a T7 promoter, suggesting that all products were translated from a single message. Expression of related products in mycoplasmas was examined by immunoblot analysis of M. hyorhinis proteins with antiserum against overexpressed recombinant proteins. A single 115-kilodalton mycoplasma protein was detected, which is larger than any of the related proteins expressed in E. coli. Our analysis indicated that translation initiation sites are used in E. coli that are not active in mycoplasmas, thereby defining differences between the translational regulatory signals of mycoplasmas and eubacteria.
author Notarnicola, S M
McIntosh, M A
Wise, K S
author_facet Notarnicola, S M
McIntosh, M A
Wise, K S
author_sort Notarnicola, S M
title Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
title_short Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
title_full Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
title_fullStr Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
title_full_unstemmed Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli.
title_sort multiple translational products from a mycoplasma hyorhinis gene expressed in escherichia coli.
publishDate 1990
url https://ncbi.nlm.nih.gov/pmc/articles/PMC209098/
https://ncbi.nlm.nih.gov/pubmed/2188946
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