Abstract
Originalsprog | Engelsk |
---|---|
Tidsskrift | Journal of Molecular Biology |
Vol/bind | 207 |
Udgave nummer | 2 |
Sider (fra-til) | 365-77 |
Antal sider | 12 |
ISSN | 0022-2836 |
DOI | |
Status | Udgivet - 1989 |
Bibliografisk note
Keywords: Bacterial Proteins; Base Sequence; Codon; Escherichia coli; Lac Operon; Molecular Sequence Data; Nucleic Acid Conformation; Protein Biosynthesis; RNA, Bacterial; RNA, Messenger; Time FactorsAdgang til dokumentet
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Codon usage determines translation rate in Escherichia coli. / Sørensen, Michael Askvad; Kurland, C G; Pedersen, Steen.
I: Journal of Molecular Biology, Bind 207, Nr. 2, 1989, s. 365-77.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review
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TY - JOUR
T1 - Codon usage determines translation rate in Escherichia coli
AU - Sørensen, Michael Askvad
AU - Kurland, C G
AU - Pedersen, Steen
N1 - Keywords: Bacterial Proteins; Base Sequence; Codon; Escherichia coli; Lac Operon; Molecular Sequence Data; Nucleic Acid Conformation; Protein Biosynthesis; RNA, Bacterial; RNA, Messenger; Time Factors
PY - 1989
Y1 - 1989
N2 - We wish to determine whether differences in translation rate are correlated with differences in codon usage or with differences in mRNA secondary structure. We therefore inserted a small DNA fragment in the lacZ gene either directly or flanked by a few frame-shifting bases, leaving the reading frame of the lacZ gene unchanged. The fragment was chosen to have "infrequent" codons in one reading frame and "common" codons in the other. The insert in these constructs does not seem to give mRNAs that are able to form extensive secondary structures. The translation time for these modified lacZ mRNAs was measured with a reproducibility better than plus or minus one second. We found that the mRNA with infrequent codons inserted has an approximately three-seconds longer translation time than the one with common codons. In another set of experiments we constructed two almost identical lacZ genes in which the lacZ mRNAs have the potential to generate stem structures with stabilities of about -75 kcal/mol. In this way we could investigate the influence of mRNA structure on translation rate. This type of modified gene was generated in two reading frames with either common or infrequent codons similar to our first experiments. We find that the yield of protein from these mRNAs is reduced, probably due to the action in vivo of an RNase. Nevertheless, the data do not indicate that there is any effect of mRNA secondary structure on translation rate. In contrast, our data persuade us that there is a difference in translation rate between infrequent codons and common codons that is of the order of sixfold.
AB - We wish to determine whether differences in translation rate are correlated with differences in codon usage or with differences in mRNA secondary structure. We therefore inserted a small DNA fragment in the lacZ gene either directly or flanked by a few frame-shifting bases, leaving the reading frame of the lacZ gene unchanged. The fragment was chosen to have "infrequent" codons in one reading frame and "common" codons in the other. The insert in these constructs does not seem to give mRNAs that are able to form extensive secondary structures. The translation time for these modified lacZ mRNAs was measured with a reproducibility better than plus or minus one second. We found that the mRNA with infrequent codons inserted has an approximately three-seconds longer translation time than the one with common codons. In another set of experiments we constructed two almost identical lacZ genes in which the lacZ mRNAs have the potential to generate stem structures with stabilities of about -75 kcal/mol. In this way we could investigate the influence of mRNA structure on translation rate. This type of modified gene was generated in two reading frames with either common or infrequent codons similar to our first experiments. We find that the yield of protein from these mRNAs is reduced, probably due to the action in vivo of an RNase. Nevertheless, the data do not indicate that there is any effect of mRNA secondary structure on translation rate. In contrast, our data persuade us that there is a difference in translation rate between infrequent codons and common codons that is of the order of sixfold.
U2 - doi:10.1016/0022-2836(89)90260-X
DO - doi:10.1016/0022-2836(89)90260-X
M3 - Journal article
C2 - 2474074
VL - 207
SP - 365
EP - 377
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 2
ER -