rnr Gene from the Antarctic Bacterium Pseudomonas syringae Lz4W, Encoding a Psychrophilic RNase R
In: Applied and Environmental Microbiology, Jg. 77 (2011-11-15), S. 7896-7904
Online
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Zugriff:
RNase R is a highly processive, hydrolytic 3′-5′ exoribonuclease belonging to the RNB/RNR superfamily which plays significant roles in RNA metabolism in bacteria. The enzyme was observed to be essential for growth of the psychrophilic Antarctic bacterium Pseudomonas syringae Lz4W at a low temperature. We present results here pertaining to the biochemical properties of RNase R and the RNase R-encoding gene ( rnr ) locus from this bacterium. By cloning and expressing a His 6 -tagged form of the P. syringae RNase R (RNase R Ps ), we show that the enzyme is active at 0 to 4°C but exhibits optimum activity at ∼25°C. The enzyme is heat labile in nature, losing activity upon incubation at 37°C and above, a hallmark of many psychrophilic enzymes. The enzyme requires divalent cations (Mg 2+ and Mn 2+ ) for activity, and the activity is higher in 50 to 150 mM KCl when it largely remains as a monomer. On synthetic substrates, RNase R Ps exhibited maximum activity on poly(A) and poly(U) in preference over poly(G) and poly(C). The enzyme also degraded structured malE-malF RNA substrates. Analysis of the cleavage products shows that the enzyme, apart from releasing 5′-nucleotide monophosphates by the processive exoribonuclease activity, produces four-nucleotide end products, as opposed to two-nucleotide products, of RNA chain by Escherichia coli RNase R. Interestingly, three ribonucleotides (ATP, GTP, and CTP) inhibited the activity of RNase R Ps in vitro . The ability of the nonhydrolyzable ATP-γS to inhibit RNase R Ps activity suggests that nucleotide hydrolysis is not required for inhibition. This is the first report on the biochemical property of a psychrophilic RNase R from any bacterium.
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rnr Gene from the Antarctic Bacterium Pseudomonas syringae Lz4W, Encoding a Psychrophilic RNase R
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Autor/in / Beteiligte Person: | Ray, M. K. ; Sulthana, Shaheen ; Rajyaguru, Purusharth I. ; Mittal, Pragya |
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Zeitschrift: | Applied and Environmental Microbiology, Jg. 77 (2011-11-15), S. 7896-7904 |
Veröffentlichung: | American Society for Microbiology, 2011 |
Medientyp: | unknown |
ISSN: | 1098-5336 (print) ; 0099-2240 (print) |
DOI: | 10.1128/aem.05683-11 |
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