Inhibition of translation by consecutive rare leucine codons in E. coli: absence of effect of varying mRNA stability.
Consecutive homologous codons that are rarely used in E. coli are known to inhibit translation to varying degrees. As few as two consecutive rare arginine codons exhibit a profound inhibition of translation when they are located in the 5' portion of a gene in E. coli. We have previously shown that nine consecutive rare CUA leucine codons cause almost complete inhibition of translation when they are placed after the 13th codon of a test message (although they do not inhibit translation when they are placed in the middle of the message). In the present work, we report that five consecutive rare CUA leucine codons exhibit approximately a threefold inhibition of translation when they are similarly placed after the 13th codon of a test message, compared to five consecutive common CUG leucine codons, in a T7 RNA polymerase-driven system. Further, by removing RNase III processing sites at the 3' ends of the mRNAs, we have manipulated the stability of the mRNAs encoding the test and control messages to see if decreasing mRNA stability might have an effect on the extent of translation inhibition by the rare leucine codons. However, the inhibition with the less stable mRNAs was similar to that with the stable mRNAs, approximately 3.4-fold, indicating that mRNA stability per se does not have a major influence on the effects of rare codons in this system.
Shu
Ping
P
Department of Microbiology & Molecular Genetics, New Jersey Medical School, University of Medicine & Dentistry of New Jersey, Newark, NJ 07103, USA.
Dai
Huacheng
H
Gao
Wenwu
W
Goldman
Emanuel
E
D020413:3' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D007930:Leucine / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping
D020413:3' Untranslated Regions; D003062:Codon; D012329:RNA, Bacterial; D012333:RNA, Messenger; D007930:Leucine