Human DNA ligase I cDNA: Cloning and functional expression in Saccharomyces cerevisiae

Journal Article · Sat Sep 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.17.6679· OSTI ID:5381681

Barnes, D E; Kodama, Kenichi; Tomkinson, A E; Lindahl, T; Lasko, D D ^[1]; Johnston, L H ^[2]

---

1. Clare Hall Labs., Herts (England)
2. National Institute for Medical Research, London (England)

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 Saccharomuces 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.

• OSTI ID: 5381681
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 87:17; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA HELICASES
AMINO ACID SEQUENCE
DNA-CLONING
LIGASES
DNA HYBRIDIZATION
DNA SEQUENCING
GENETIC MAPPING
HUMAN CHROMOSOME 19
MAN
SACCHAROMYCES CEREVISIAE
ANIMALS
CHROMOSOMES
CLONING
ENZYMES
EUMYCOTA
FUNGI
HYBRIDIZATION
MAMMALS
MAPPING
MICROORGANISMS
MOLECULAR STRUCTURE
PLANTS
PRIMATES
SACCHAROMYCES
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
YEASTS
550201* - Biochemistry- Tracer Techniques
550401 - Genetics- Tracer Techniques