Reactivation of XIST in normal fibroblasts and a somatic cell hybrid: abnormal localization of XIST RNA in hybrid cells.
The XIST gene, expressed only from the inactive X chromosome, is a critical component of X inactivation. Although apparently unnecessary for maintenance of inactivation, XIST expression is thought to be sufficient for inactivation of genes in cis even when XIST is located abnormally on another chromosome. This repression appears to involve the association of XIST RNA with the chromosome from which it is expressed. Reactivated genes on the inactive X chromosome, however, maintain expression in several somatic cell hybrid lines with stable expression of XIST. We describe here another example of an XIST-expressing human-hamster hybrid that lacks X-linked gene repression in which the human XIST gene present on an active X chromosome was reactivated by treatment with 5-aza-2'-deoxycytidine. These data raise the possibility that human XIST RNA does not function properly in human-rodent somatic cell hybrids. As part of our approach to address this question, we reactivated the XIST gene in normal male fibroblasts and then compared their patterns of XIST RNA localization by subcellular fractionation and in situ hybridization with those of hybrid cells. Although XIST RNA is nuclear in all cell types, we found that the in situ signals are much more diffuse in hybrids than in human cells. These data suggest that hybrids lack components needed for XIST localization and, presumably, XIST-mediated gene repression.
Hansen
R S
RS
Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Canfield
T K
TK
Stanek
A M
AM
Keitges
E A
EA
Gartler
S M
SM
D000071443:12E7 Antigen; D000818:Animals; D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D002467:Cell Nucleus / Q000378:metabolism; D006224:Cricetinae; D005260:Female; D005347:Fibroblasts; D005786:Gene Expression Regulation; D006822:Hybrid Cells / Q000502:physiology*; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D062085:RNA, Long Noncoding; D012333:RNA, Messenger / Q000235:genetics; D022661:RNA, Untranslated; D014157:Transcription Factors / Q000502:physiology*; D014960:X Chromosome
D000071443:12E7 Antigen; D015703:Antigens, CD; C061828:CD99 protein, human; D015815:Cell Adhesion Molecules; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D022661:RNA, Untranslated; D014157:Transcription Factors; C077885:XIST non-coding RNA