On the localization of ribonucleases in bacteria.
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Biomedical subjects
Publications and source records attributed to D Apirion.
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The question of whether or not a cellular ribonuclease is involved in the cleavage of 16S ribosomal RNA by colicin E3 was investigated. For this purpose ribosomes from strains devoid of some ribonucleases or ribosomes in which ribonucleases had been inactivated by heat or removed by extensive washings were used for the colicin reaction. Since the 16S RNA of all these different ribosomes, and even of the most extensively washed ribosomes, was cleaved by colicin E3, it is suggested that cellular ribonucleases are not involved in colicin E3 action. Thus, colicin E3 seems to be a unique endoribonuclease.
Mutants having low levels of polynucleotide phosphorylase activity grow poorly at 45 C. All revertants isolated for their ability to grow better at that temperature also regained higher levels of polynucleotide phosphorylase and the ability to be induced for tryptophanase. Thus, a physiological role is implied for the enzyme polynculeotide phosphorylase.
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3',5' cyclic-AMP (cAMP) will stimulate the rate of tryptophanase synthesis in Escherichia coli cultures induced with tryptophan. Adding cAMP after the initiation of messenger RNA synthesis was blocked by rifampicin, did not stimulate tryptophanase synthesis. This indicates that cAMP acts at initiation of either transcription or translation and not at the level of chain elongation of either the messenger or the polypeptide chain.
In a temperature-sensitive mutant of Escherichia coli, beta-galactosidase cannot be induced at the nonpermissive temperature (43 degrees C) without the addition of exogenous 3', 5'-cyclic AMP (cAMP), although the intracellular concentration of this nucleotide is normal. This specific effect of cAMP is probably general in this strain for those operons which are controlled by the cAMP receptor protein and cAMP, but not for other parts of the chromosome. The lac mRNA produced at 43 degrees C in absence of cAMP is transcribed from the correct DNA strand and it directs the synthesis of enzymatically inactive material cross-reacting with beta-galactosidase. Experiments separating transcription from translation by using rifampicin, suggest that cAMP exerts its effect during initiation of transcription or translation of lac mRNA, but does not affect the propagation of either the messenger or of the beta-galactosidase polypeptide chain.
A mutant strain of Escherichia coli previously thought to possess low levels of ribonuclease II activity has normal levels of ribonuclease II after partial purification of this enzyme from crude extracts.
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A temperature-sensitive mutant strain of Escherichia coli exhibits a remarkable increase in RNase activity when grown at its nonpermissive temperature. During growth at the nonpermissive temperature, there is an increase in the extent of breakdown of pulse-labeled RNA and a decrease in the functional lifetime of the mRNA for the lac operon. T7 RNA, which is usually stable in E. coli, is also stable in this strain at the nonpermissive temperature. It is possible that the RNase measured is part of the normal mechanism of mRNA degradation in the cell. A mechanism for mRNA degradation that requires the combined action of endonuclease(s) and 3' to 5' exonuclease(s) is proposed.
Aurintricarboxylic acid (ATA) inhibits protein synthesis directed by polyuridylic acid. All steps tested are inhibited by ATA. We conclude that inhibition of polyphenylalanine formation is an additive effect of inhibition of various steps in the protein synthetic machinery.
The effect of aurintricarboxylic acid (ATA) was tested on various aspects of protein synthesis directed by the natural messenger ribonucleic acid (RNA) isolated from R17 RNA bacteriophage. The effects of various levels of ATA (up to 1,000 mum) were tested on overall protein synthesis as well as on binding of messenger RNA and fmet-transfer RNA to ribosomes and on the addition of the 50S ribosome to the 30S ribosome initiation complex. All of the reactions tested could be inhibited by ATA, and none of the tested steps was found to be uniquely sensitive to it. However, the total initiation steps were more sensitive to this chemical than the elongation steps; thus, under appropriate conditions this chemical can preferentially inhibit initiation while elongation of the polypeptide chain is not appreciably affected.