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J Biol Chem, Vol. 273, Issue 31, 19895-19901, July 31, 1998
From the ** Howard Hughes Medical Institute and Departments of
A human MSH2-human MSH3 (hMSH2·hMSH3) complex
of approximately 1:1 stoichiometry (human MutS
Isolation of MutS
from Human Cells and Comparison of the
Mismatch Repair Specificities of MutS and MutS
,¶,, and**
Biochemistry and ¶ Medicine-Medical Oncology, Duke
University Medical Center, Durham, North Carolina 27710
(hMutS)) has been
demonstrated in several human tumor cell lines and purified to near
homogeneity. In vitro, hMutS supports the efficient
repair of insertion/deletion (I/D) heterologies of 2-8 nucleotides, is
weakly active on a single-nucleotide I/D mispair, and is not detectably
active on the eight base-base mismatches. Human MutS
(hMutS), a
heterodimer of hMSH2 and hMSH6, efficiently supports the repair of
single-nucleotide I/D mismatches, base-base mispairs, and all
substrates tested that were repaired by hMutS. Thus, the repair
specificities of hMutS and hMutS are redundant with respect to
the repair of I/D heterologies of 2-8 nucleotides. The hMutS level
in repair-proficient HeLa cells (1.5 µg/mg nuclear extract) is
approximately 10 times that of hMutS. In HCT-15 colorectal tumor
cells, which do not contain hMSH6 and consequently lack hMutS, the
hMutS level is elevated severalfold relative to that in HeLa cells
and is responsible for the repair of I/D mismatches that has been
observed in this cell line. LoVo tumor cells, which are genetically
deficient in hMSH2, lack both hMutS and hMutS, and hMSH3 and
hMSH6 levels are less than 4% of those found in repair-proficient
cells. Coupled with previous findings (J. T. Drummond, J. Genschel, E. Wolf, and P. Modrich (1997) Proc. Natl. Acad. Sci.
U. S. A. 94, 10144-10149), these results suggest that hMSH2
partitions between available pools of hMSH3 and hMSH6 and indicate that
hMSH2 positively modulates hMSH6 and hMSH3 levels, perhaps by
stabilization of the polypeptides upon heterodimer formation.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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