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PMID:9931460

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Citation

Chamankhah, M, Wei, YF and Xiao, W (1998) Isolation of hMRE11B: failure to complement yeast mre11 defects due to species-specific protein interactions. Gene 225:107-16

Abstract

The Saccharomyces cerevisiae MRE11 gene plays an important role in meiotic recombination, mitotic DNA repair and telomere maintenance. We present the isolation of hMRE11B cDNA from a human HeLa cell cDNA library as an MRE11 homolog. Compared to the previously identified hMRE11, hMRE11B contains an additional 84bp sequence that results in a 28 amino-acid insertion close to the C-terminus. The expression pattern of hMRE11B in different tissues shows the presence of two mRNA species of approx. 2.6 and 7.5kb. Overexpression of hMRE11B does not complement the alkylation sensitivity of the mre11 null and temperature-sensitive mutant strains. In this study, we examine factors that may explain this lack of complementation. First, both Northern and Western analyses rule out the lack of hMRE11B transcription and/or translation in yeast. Second, we demonstrate that hMre11B, like the yeast Mre11 protein, dimerizes in vivo in a yeast two-hybrid system. This dimerization requires the C-terminal one-third of hMre11B protein, which includes the 28 amino acids absent in hMre11. However, hMre11B does not interact with Mre11, Rad50 and Xrs2. Hence, the lack of protein-protein interaction between hMre11B and the yeast Mre11, Rad50, and Xrs2 may explain the inability of hMRE11B to complement the yeast mre11 mutants. We rule out the hypothesis that the lack of interaction and, in turn of complementation, is due to the absence of sequence homology at the C-terminal domain of hMre11B compared to the yeast Mre11. Instead, we propose that the C-terminus of hMre11B participates in protein-protein interaction and functions in a species-specific manner.

Links

PubMed

Keywords

Base Sequence; Binding Sites; Blotting, Northern; Cloning, Molecular; DNA, Complementary/chemistry; DNA, Complementary/genetics; DNA, Complementary/isolation & purification; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Dimerization; Endodeoxyribonucleases; Exodeoxyribonucleases; Fungal Proteins/genetics; Fungal Proteins/metabolism; Gene Expression; Genetic Complementation Test; HL-60 Cells/cytology; HL-60 Cells/metabolism; HeLa Cells; Humans; K562 Cells/cytology; K562 Cells/metabolism; Molecular Sequence Data; Mutation; RNA/genetics; RNA/metabolism; Recombinant Fusion Proteins/genetics; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Species Specificity; Tissue Distribution; Tumor Cells, Cultured/cytology; Tumor Cells, Cultured/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:MRE11

involved_in

GO:0007131: reciprocal meiotic recombination

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:MRE11

involved_in

GO:0007004: telomere maintenance via telomerase

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:MRE11

involved_in

GO:0006281: DNA repair

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:MRE11

involved_in

GO:0007131: reciprocal meiotic recombination

ECO:0000304: author statement supported by traceable reference used in manual assertion


P

Seeded From UniProt

complete

HUMAN:MRE11

involved_in

GO:0006281: DNA repair

ECO:0000304: author statement supported by traceable reference used in manual assertion


P

Seeded From UniProt

complete


See also

References

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