[Preparation and characterization of nuclear matrix/chromosome scaffold in situ].
A method of nuclear matrix and chromosomal scaffold preparation from cultured animal cells was developed. After the high-salt extraction, interphase and mitotic cells were not detached from the coverslips that enabled us to analyse the nuclear matrix and chromosomal scaffold in cells at all mitotic phases. Morphological methods (phase contrast microscopy and electron microscopy of ultrathin sections) did not reveal any structures that could be identified as a chromosomal scaffold. However, after staining with antibodies to XCAP-E and topoisomerase IIalpha some structures were revealed in metaphase cells having both localization and morphology of a chromosomal scaffold. The cell residuals were not stained with antibodies to XCAP-E and topoisomerase IIalpha, if the nuclear matrix and chromosomal scaffold were destabilized by addition of beta-mercaptoethanol.
Sheval'
E V
EV
Kurchashova
S Iu
SIu
Timirbulatova
E R
ER
Poliakov
V Iu
VIu
D000818:Animals; D000951:Antigens, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism; D018797:Cell Cycle Proteins; D002875:Chromosomes / Q000378:metabolism* / Q000648:ultrastructure; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D045170:Matrix Attachment Regions; D051379:Mice; D008854:Microscopy, Electron; D015530:Nuclear Matrix / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D012965:Sodium Chloride / Q000737:chemistry; D013552:Swine; D016707:Tissue Fixation / Q000379:methods*; D029867:Xenopus Proteins / Q000302:isolation & purification / Q000378:metabolism; D014982:Xenopus laevis
D000951:Antigens, Neoplasm; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C090453:XCAP-E protein, Xenopus; D029867:Xenopus Proteins; D012965:Sodium Chloride; D004250:DNA Topoisomerases, Type II