[Endogenous DNases as a tool for isolation of nuclear matrix: critical parameters of nucleolysis].
Degree of nucleolysis has critical significance for isolation of nuclear matrix (NM) specifically enriched in transcribed DNA sequences as demonstrated at the example of inactive (c-fos, c-myc, and Ck) and active (p53, albumin, and 28S rRNA) genes in resting hepatocytes. Optimal degree of nucleolysis features degradation of loop domains of chromatin with preserved relatively uniform molecular weight distribution of DNA. Deviation from these parameters leads to nonspecific fragmentation of chromatin in various gene loci and isolation of NM samples nonspecifically enriched or depleted of transcribed DNA sequences. Under optimal hydrolytic conditions, the transcribed chromatin is more resistant to endogenous DNase attack, which allows selective conservation of its association with the nuclear matrix.
Borisova
N P
NP
Institute of Problems of Chemical Physics, Russian Academy of Sciences, prosp. Akad. Semenova 4, Chernogolovka, Moscow Region, 142432 Russia.
Kostiuk
G V
GV
Shevchenko
N A
NA
Boĭkov
P Ia
PIa
Rudakova
E V
EV
Papina
R I
RI
Terent'ev
A A
AA
D000418:Albumins / Q000235:genetics; D000818:Animals; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D015870:Gene Expression; D016158:Genes, p53 / Q000235:genetics; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008297:Male; D015530:Nuclear Matrix / Q000235:genetics / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D011519:Proto-Oncogenes / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D014158:Transcription, Genetic
D000418:Albumins; D002843:Chromatin; D012339:RNA, Ribosomal, 28S; D004247:DNA; D003851:Deoxyribonucleases