Characterization of RNase P from Thermotoga maritima.
The protein subunit of RNase P from a thermophilic bacterium, Thermotoga maritima, was overexpressed in and purified from Escherichia coli. The cloned protein was reconstituted with the RNA subunit transcribed in vitro. The temperature optimum of the holoenzyme is near 50 degrees C, with no enzymatic activity at 65 degrees C or above. This finding is in sharp contrast to the optimal growth temperature of T.maritima, which is near 80 degrees C. However, in heterologous reconstitution experiments in vitro with RNase P subunits from other species, we found that the protein subunit from T.maritima was responsible for the comparative thermal stability of such complexes.
Paul
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Department of Molecular, Cellular and Developmental Biology, Yale University, PO Box 208103, New Haven, CT 06520-8103, USA.
Lazarev
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Altman
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D004722:Endoribonucleases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D020035:Holoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D021122:Protein Subunits; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D012365:RNA, Transfer, Tyr / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D043262:Ribonuclease P; D016415:Sequence Alignment; D013696:Temperature; D020124:Thermotoga maritima / Q000201:enzymology* / Q000235:genetics*; D014158:Transcription, Genetic
D000596:Amino Acids; D029968:Escherichia coli Proteins; D020035:Holoenzymes; D021122:Protein Subunits; D012329:RNA, Bacterial; D016337:RNA, Catalytic; D012365:RNA, Transfer, Tyr; D011993:Recombinant Fusion Proteins; D004722:Endoribonucleases; D043262:Ribonuclease P; C470865:ribonuclease P, E coli