title: Transcription- and apoptosis-dependent long-range distribution of tight DNA-protein complexes in the chicken alpha-globin gene.
abstract: The proteins tightly bound to DNA (TBP) are a group of proteins that remain attached to DNA with covalent or noncovalent bonds after its deproteinization, and have been hypothesized to be involved in regulation of gene expression. To investigate this question further, oligonucleotide DNA arrays were used to determine the distribution of tightly bound proteins along a 100-kb DNA fragment surrounding the chicken alpha-globin gene domain in DNA from chicken erythrocytes, liver, and AEV-transformed HD3 (erythroblast) cells in different physiological conditions. DNA was fractionated into TBP-free (F) and TBP-enriched (R) fractions by separation on nitrocellulose, and these fractions were used as probes for hybridization with the microarray. In erythrocytes, the site 60 kb from the 5' end of the sequence and containing a LINE family CR1 repeat was TBP enriched, but in HD3 cells this sequence was devoid of TBPs. Thus cessation of transcription of the domain is followed by an F-R transition of this site. In apoptotic HD3 cells, TBPs remained attached to DNA only at a site situated 16 kb from the 5' end of the sequence. These data confirm and extend previous conclusions about the specificity of the DNA sequences that preferably form tight complexes with proteins and about the differentiation-specific distribution of the TBPs in different cell lineages. Binding of TBPs appears to be independent of primary DNA sequence.
authors.affiliation: Department of Biochemistry and Biophysics, Vilnius University, Vilnius, Lithuania.
authors.forename: Kristina
authors.initials: K
authors.lastname: Bielskiene
authors.forename: Lida
authors.initials: L
authors.lastname: Bagdoniene
authors.forename: Benediktas
authors.initials: B
authors.lastname: Juodka
authors.forename: Marc
authors.initials: M
authors.lastname: Lipinski
authors.forename: Tatjana
authors.initials: T
authors.lastname: Sjakste
authors.forename: Yegor S
authors.initials: YS
authors.lastname: Vassetzky
authors.forename: Nikolajs
authors.initials: N
authors.lastname: Sjakste
mesh_terms: D000818:Animals; D017209:Apoptosis; D002645:Chickens / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D005914:Globins / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D005914:Globins