Cloning of a cDNA from Arabidopsis thaliana homologous to the human XPB gene.
The human gene XPB, defective in xeroderma pigmentosum patients complementation group B, encodes a DNA helicase involved in several DNA metabolic pathways, including DNA repair and transcription. The high conservation of this gene has allowed the cloning of homologs in various species, such as mouse, yeast and Drosophila. Not much information on the molecular basis of nucleotide excision repair in plants is available, but these organisms may have similar mechanisms to other eukaryotes. A homolog of XPB was isolated in Arabidopsis thaliana by using polymerase chain reaction (PCR) with degenerate oligonucleotides based on protein domains which are conserved among several species. Screening of an Arabidopsis cDNA library led to the identification and isolation of a cDNA clone with 2670 bp encoding a predicted protein of 767 amino acids, denoted araXPB. Genomic analysis indicated that this is a nuclear single copy gene in plant cells. Northern blot with the cDNA probe revealed a major transcript which migrated at approx. 2,800 b, in agreement with the size of the cDNA isolated. The araXPB protein shares approximately 50% identical and 70% conserved amino acids with the yeast and human homologs. The plant protein maintains all the functional domains found in the other proteins, including nuclear localization signal, DNA-binding domain and helicase motifs, suggesting that it might also act as part of the RNA transcription apparatus, as well as nucleotide excision repair in plant cells.
Ribeiro
D T
DT
Department of Biology, University of São Paulo, Brazil.
Machado
C R
CR
Costa
R M
RM
Praekelt
U M
UM
Van Sluys
M A
MA
Menck
C F
CF
D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics*; D018076:DNA, Complementary; D018744:DNA, Plant / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D004330:Drosophila / Q000235:genetics; D017343:Genes, Plant; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014983:Xeroderma Pigmentosum / Q000235:genetics
D018076:DNA, Complementary; D018744:DNA, Plant; D004268:DNA-Binding Proteins; C072022:XPBC-ERCC-3 protein; D004265:DNA Helicases