J. Biol. Chem., Vol. 267, Issue 25, 17531-17539, Sep, 1992
DNA polymerase-delta from the silk glands of Bombyx mori
S Niranjanakumari and KP Gopinathan
Microbiology and Cell Biology Department, Indian Institute of Science, Bangalore.
The silk gland of Bombyx mori is a terminally differentiated tissue in
which DNA replication continues without cell or nuclear division during
larval development. DNA polymerase-delta activity increases in the
posterior and middle silk glands during the development period, reaching
maximal levels in the middle of the fifth instar larvae. The enzyme has
been purified to homogeneity by a series of column chromatographic and
affinity purification steps. It is a multimer comprising of three
heterogeneous subunits, M(r) 170,000, 70,000, and 42,000. An auxiliary
protein from B. mori silk glands, analogous to the proliferating cell
nuclear antigen, enhances the processivity of the enzyme and stimulates
catalytic activity by 3-fold. This auxiliary protein has also been purified
to homogeneity. It is a dimer comprised of a single type M(r) 40,000
subunit. Polymerase-delta possesses an intrinsic 3'----5' exonuclease
activity which participates in proofreading by mismatch repair during DNA
synthesis and is devoid of any primase activity. DNA polymerase-delta
activity could be further distinguished from polymerase-alpha from the same
tissue based on its sensitivity to various inhibitors and polyclonal
antibodies to the individual enzymes. Like DNA polymerase-alpha,
polymerase-delta is also tightly associated with the nuclear matrix. The
polymerase alpha- primase complex could be readily separated from
polymerase-delta (exonuclease) in the purification protocol adopted. DNA
polymerase- delta from B. mori silk glands resembles the mammalian delta-
polymerases. Considering that both DNA polymerase-delta and -alpha are
present in nearly equal amounts in this highly replicative tissue and their
close association with the nuclear matrix, the involvement of both the
enzymes in the chromosomal endoreplication process in B. mori is strongly
implicated.