Abstract
The human SNM1 protein is a member of a highly conserved group of proteins catalyzing the hydrolysis of nucleic acid substrates. Although overproduction is unstable in mammalian cells, we have overproduced a recombinant hSNM1 protein in an insect cell system. The protein is a single-strand 5'-exonuclease, like its yeast homolog. The enzyme utilizes either DNA or RNA substrates, requires a 5'-phosphate moiety, shows very little activity on double-strand substrates, and functions at a size consistent with a monomer. The exonuclease activity requires the conserved beta-lactamase domain; site-directed mutagenesis of a conserved aspartate inactivates the exonuclease.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Animals
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Aspartic Acid / genetics
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Cell Line
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DNA Repair Enzymes / chemistry
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DNA Repair Enzymes / genetics
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DNA Repair Enzymes / metabolism*
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DNA, Single-Stranded / metabolism
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Drosophila melanogaster / cytology
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Endodeoxyribonucleases / metabolism
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Exodeoxyribonucleases / chemistry
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Exodeoxyribonucleases / genetics
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Exodeoxyribonucleases / metabolism*
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Fibroblasts / metabolism
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Humans
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Mutagenesis, Site-Directed
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Nuclear Proteins / chemistry
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Phosphates / metabolism
Substances
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DNA, Single-Stranded
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Nuclear Proteins
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Phosphates
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Aspartic Acid
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DCLRE1B protein, human
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Endodeoxyribonucleases
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Exodeoxyribonucleases
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DNA Repair Enzymes