The 33 kb transcript of the chicken alpha-globin gene domain is part of the nuclear matrix.
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 alpha-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.
Razin
Sergey V
SV
Institut J Monod, 2, Place Jussieu, 75251 Paris, Cedex 05, France. sergey.v.razin@usa.net
Rynditch
Alla
A
Borunova
Victoria
V
Ioudinkova
Elena
E
Smalko
Victor
V
Scherrer
Klaus
K
D000818:Animals; D015152:Blotting, Northern; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002645:Chickens / Q000235:genetics*; D041905:Erythroid Cells / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D005914:Globins / Q000235:genetics*; D017403:In Situ Hybridization; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics; D015530:Nuclear Matrix / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
D012333:RNA, Messenger; D005914:Globins