Qp. Cao et al., THE 68-KDA CALMODULIN-BINDING PROTEIN IS TIGHTLY ASSOCIATED WITH THE MULTIPROTEIN DNA-POLYMERASE ALPHA-PRIMASE COMPLEX IN HELA-CELLS, Biochemistry, 34(12), 1995, pp. 3878-3883
Calcium and its receptor protein calmodulin function in the regulation
of proliferation of mammalian cells. A 68 kDa calmodulin-specific bin
ding protein was shown previously to be associated with growth factor-
dependent progression of a variety of mammalian cells from G1 to S pha
se and to stimulate DNA synthesis in permeabilized hematopoietic proge
nitor cells, In this report we show that the 68 kDa calmodulin-specifi
c binding protein in HeLa cells is tightly associated with the DNA pol
ymerase a-primase component of the 21S complex of enzymes for DNA synt
hesis. The 68 kDa calmodulin-binding protein and the DNA polymerase cr
-primase complex cofractionate during Q-Sepharose chromatography to is
olate the 21S enzyme complex, native and denatured DNA-cellulose to di
ssociate the 21S complex, and DEAE-Bio-Gel chromatography to isolate t
he multiprotein DNA polymerase cc-primase complex. The 68 kDa calmodul
in-specific binding protein and DNA polymerase or also bind and coelut
e during affinity chromatography on calmodulin-agarose. They also copr
ecipitate with C10-agarose-linked monoclonal antibody SJK 132-20 to hu
man DNA polymerase or. The tight association of the 68 kDa calmodulin-
binding protein to the DNA polymerase alpha-primase complex supports a
function for this protein in the regulation of DNA synthesis in vivo.