Open AccessDNA regions associated with the nuclear matrix of Ehrlich ascites cells expose single‐stranded sites after deproteinization
Author(s) -
PROBST Hans,
HERZOG Richard
Publication year - 1985
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1985.tb08634.x
Subject(s) - nuclease , dna , nuclear matrix , nitrocellulose , endonuclease , thymidine , biochemistry , restriction enzyme , matrix (chemical analysis) , microbiology and biotechnology , chemistry , biology , chromatography , membrane , chromatin
Ehrlich ascites cells were pulse‐labeled with [ 3 H]thymidine and subjected to prolonged labeling with [ 14 C]thymidine. The isolated nuclei were digested with the restriction endonuclease Bsp RI and then processed to yield a ‘matrix fraction’ and a ‘non‐matrix fraction’. The DNA fragments purified from these fractions and from whole digested nuclei were examined for nitrocellulose‐binding sites before and after digestion with single‐strand‐specific (S1) nuclease. Both, pulse‐labeled and long‐time‐labeled fragments, isolated from the matrix fraction, exhibited a significantly increased content of nitrocellulose‐binding sites. The major portion of these sites were rendered non‐binding by digestion with single‐strand‐specific nuclease and consisted most probably of structures exposing relatively small stretches of non‐base‐paired DNA. The nature of the minor portion of binding sites which was insensitive to single‐strand‐specific nuclease is not clear. Both types of binding sites are possible candidates for mediating the attachment of DNA to the nuclear matrix.