Human DNA ligase I cDNA: cloning and functional expression in Saccharomyces cerevisiae.
Human cDNA clones encoding the major DNA ligase activity in proliferating cells, DNA ligase I, were isolated by two independent methods. In one approach, a human cDNA library was screened by hybridization with oligonucleotides deduced from partial amino acid sequence of purified bovine DNA ligase I. In an alternative approach, a human cDNA library was screened for functional expression of a polypeptide able to complement a cdc9 temperature-sensitive DNA ligase mutant of Saccharomyces cerevisiae. The sequence of an apparently full-length cDNA encodes a 102-kDa protein, indistinguishable in size from authentic human DNA ligase I. The deduced amino acid sequence of the human DNA ligase I cDNA is 40% homologous to the smaller DNA ligases of S. cerevisiae and Schizosaccharomyces pombe, homology being confined to the carboxyl-terminal regions of the respective proteins. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is transcribed from a single-copy gene on chromosome 19.
Barnes
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DE
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, United Kingdom.
Johnston
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LH
Kodama
K
K
Tomkinson
A E
AE
Lasko
D D
DD
Lindahl
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T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000096:biosynthesis / Q000235:genetics*; D005816:Genetic Complementation Test; D006801:Humans; D006822:Hybrid Cells / Q000201:enzymology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011118:Polynucleotide Ligases / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
C000607890:CDC9 protein, S cerevisiae; C000607892:LIG1 protein, human; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D011088:DNA Ligases; D011118:Polynucleotide Ligases; D000072481:DNA Ligase ATP