The Effects of Anti-tumor Antibiotic CC-1065 on Restriction Enzymes Involved in Processing the DNA
Loading...
Authors
Iglesias, Norma-Irene
Issue Date
1982
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
CC-1065 is a potent new anti-tumor antibiotic discovered
in Upjohn Laboratories, produced by Streptomyces zelensis.
This drug interacts strongly with double stranded DNA at
adenine-thymine sites in the minor groove and is thought to
exert its cytotoxic effects through superstabilization of the
DNA helix. The manner in which it binds to DNA suggests
possible mechanisms of its cytotoxicity such as stiffening of
the helix by CC-1065 which prevents free association of
the DNA with the enzyme of proper enzyme activity, physical
masking of an enzyme restriction site, and inhibition of normal
unwinding and enzymatic processes occurring during replication.
In this project the effects of CC-1065 on restriction
enzymes specific for known sequences on ππ²174RF phage DNA were
investigated. The ππ²174RF DNA was treated with various
concentrations of CC-1065 so as to yield CC-1065 molecule to
nucleotide ratios of 1:50, 1:200, and 1:800. After this the
samples were treated with enzyme. A control was also
prepared containing ,ππ²174RF DNA and enzyme ( no drug ), another
was prepared containing the CC-1065 and ,ππ²174RF DNA alone
( no enzyme,). These samples were incubated for 1,2, or 3
hours at 37ΒΊ C and run through agarose of polyacrylamide gel
electrophoresis. It was found that AT specific enzymes were
more greatly inhibited by CC-1065 than GC specific enzymes.
It was also found that CC-1065 does inhibit some cleavage
sites on the ππ²174RF DNA preferentially to others. Further
experimentation is necessary to determine the length of the
adenine-thymine run required by the drug, and to investigate
the long-range inductive effects of the drug.
Description
v, 35 p.
Citation
Publisher
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder. All rights reserved.