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The 33 kb transcript of the chicken α-globin gene domain is part of the nuclear matrix

✍ Scribed by Sergey V. Razin; Alla Rynditch; Victoria Borunova; Elena Ioudinkova; Victor Smalko; Klaus Scherrer


Publisher
John Wiley and Sons
Year
2004
Tongue
English
Weight
329 KB
Volume
92
Category
Article
ISSN
0730-2312

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✦ Synopsis


Abstract

Giant nuclear transcripts, and in particular the RNAs of the globin gene domains which are much larger than their canonical pre‐mRNAs, have been an enigma for many years. We show here that in avian erythroblastosis virus (AEV)‐transformed chicken erythroleukaemic cells, where globin gene expression is abortive, the whole domain of α‐globin genes is transcribed for about 33 kb in the globin direction and that this RNA is part of the nuclear matrix. Northern blot hybridisation with strand‐specific riboprobes, recognising genes and intergenic sequences, and RT‐PCR with downstream primers, show that the continuous full domain transcript (FDT) starts in the vicinity of a putative LCR and includes all the genes as well as known regulatory sites, the replication origin, and the DNA loop anchorage region in the upstream area. Absent in chicken fibroblasts, the globin FDT overlaps the major part of the ggPRX housekeeping gene that is transcribed in the opposite direction. RT‐PCR and in situ hybridisation with genic and extra‐genic globin probes demonstrated that the globin FDT is a component of the nuclear matrix. We suggest that the globin FDTs keep the domain in an active state, and the globin RNAs on the processing pathway are a component of the nuclear matrix. They may take part in the dynamic nuclear architecture when productively processed, or turn over slowly when globins are not synthesised. © 2004 Wiley‐Liss, Inc.


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We have cloned and sequenced a genomic DNA fragment of chicken containing a cluster of DNase I hypersensitive sites (DHS) located 11-15 kb upstream from the first gene of the ␣-globin gene domain and including a constitutive DHS flanked by two erythroid-specific ones. A 1.2-kb subfragment of the DNA