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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
2009-4-16
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pubmed:abstractText |
Clp1 proteins are essential components of the eukaryal mRNA 3' cleavage-polyadenylation machinery. Human Clp1 has an additional function as an RNA-specific 5'-OH polynucleotide kinase, which is implicated in RNA end healing. Yeast Clp1 has no kinase activity, although it binds ATP. Here we report that Clp1-like proteins are extant in archaea. Purification and characterization of Pyrococcus horikoshii Clp1 (PhoClp1) reveals it to be a thermostable 5'-OH polynucleotide kinase optimally active at 55 degrees C to 85 degrees C. PhoClp1 catalyzes transfer of the gamma phosphate from ATP (K (m) 16 microM) to either 5'-OH RNA or DNA ends, although it prefers RNA in a competitive situation. Increasing the monovalent salt concentration to 250 mM suppresses the DNA kinase without affecting RNA phosphorylation, suggesting that RNA is a likely substrate for this enzyme in vivo. Indeed, we show that expression of PhoClp1 in budding yeast can complement a lethal mutation in the 5'-OH RNA kinase module of tRNA ligase. PhoClp1 is a member of the P-loop phosphotransferase superfamily. Alanine mutations at the P-loop lysine (Lys49) and a conserved aspartate (Asp73) inactivate the kinase. Our studies fortify emerging evidence for an enzymatic RNA repair capacity in archaea and provide a new reagent for polynucleotide phosphorylation at high temperatures.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-10882122,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-10917597,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-11335730,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-12220496,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-12408823,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-12592006,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-14747466,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-14754987,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-14973195,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15056675,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15375211,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15579472,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15653639,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15749016,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-15987807,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-16428247,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-17151076,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-17379808,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18262407,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18511537,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18648070,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18657509,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18703739,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18829718,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-18971321,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-1922054,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-226541,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-2444436,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-4287929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-4287930,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-4366306,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-5323016,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-6186399,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-6413507,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-6828385,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19299550-9223284
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
1469-9001
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
15
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
923-31
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pubmed:dateRevised |
2010-9-22
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pubmed:meshHeading |
pubmed-meshheading:19299550-Amino Acid Sequence,
pubmed-meshheading:19299550-Humans,
pubmed-meshheading:19299550-Molecular Sequence Data,
pubmed-meshheading:19299550-Nuclear Proteins,
pubmed-meshheading:19299550-Phosphotransferases,
pubmed-meshheading:19299550-Polynucleotide 5'-Hydroxyl-Kinase,
pubmed-meshheading:19299550-Pyrococcus horikoshii,
pubmed-meshheading:19299550-RNA,
pubmed-meshheading:19299550-RNA Ligase (ATP),
pubmed-meshheading:19299550-Temperature,
pubmed-meshheading:19299550-Transcription Factors
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pubmed:year |
2009
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pubmed:articleTitle |
Characterization of a thermostable archaeal polynucleotide kinase homologous to human Clp1.
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pubmed:affiliation |
Graduate Program in Chemical Biology, Sloan-Kettering Institute, New York, New York 10065, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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