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2.
Acta Naturae ; 8(1): 90-7, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27099788

RESUMEN

A systematic search for DNA fragments containing potential CTCF transcription factor binding sites in the chicken alpha-globin domain and its flanking regions was performed by means of the two-dimension electrophoretic mobility shift assay. For the alpha-globin domain fragments selected, the occupancy by the CTCF in erythroid and lymphoid chicken cells was tested by chromatin immunoprecipitation. Only one of 13 DNA fragments capable of CTCF binding in vitro was efficiently bound to this protein in vivo in erythroid cells, and somewhat less efficiently - in lymphoid cells. So, binding of CTCF to the DNA fragment in vitro in most cases does not mean that this fragment will be occupied by CTCF in the cell nucleus. Yet, CTCF binding in vivo, as a rule, is accompanied by the binding of the protein to this DNA region in vitro. During the erythroid differentiation, no significant changes in CTCF binding to the DNA fragments studied were detected.

3.
Klin Med (Mosk) ; 94(10): 780-4, 2016.
Artículo en Ruso | MEDLINE | ID: mdl-30299043
4.
Biochemistry (Mosc) ; 78(8): 879-83, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24228875

RESUMEN

The chicken gene for transcription factor CTCF was expressed in COS-1 mammalian cells. The CTCF protein containing polyhistidine tag was partially purified using metallo-affinity and ion-exchange chromatography. The expressed protein localized in the cell nucleus and was shown to be functionally active in the electrophoretic mobility shift assay and specifically interacted with anti-CTCF antibodies.


Asunto(s)
Proteínas Represoras/aislamiento & purificación , Animales , Factor de Unión a CCCTC , Células COS , Núcleo Celular/metabolismo , Pollos , Chlorocebus aethiops , Expresión Génica , Proteínas Represoras/genética , Proteínas Represoras/metabolismo
5.
Biochemistry (Mosc) ; 76(8): 951-7, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22022969

RESUMEN

A detailed functional and evolutionary analysis of an enhancer element of the human genome (enhancer 12) located in the second intron of the U2AF1L4 gene, which we identified earlier, is presented. Overlapping fragments of the studied genome region were analyzed for enhancer activity, and the site responsible for the activity of this element was identified using transient transfections of HeLa cells. Comparison of the enhancer 12 sequence with orthologous sequences from seven primate species revealed the existence of evolutionarily conserved sequences within this element. One of the identified conservative regions is likely responsible for the enhancer activity and is able to specifically interact in vitro with proteins of HeLa cell nuclear extract. The ability of orthologous primate sequences to compete with enhancer 12 for binding with HeLa cell nuclear extract proteins and to enhance the activity of the reporter gene in transient transfection of HeLa cells is demonstrated.


Asunto(s)
Elementos de Facilitación Genéticos , Intrones , Proteínas Nucleares/genética , Ribonucleoproteínas/genética , Secuencia de Bases , Secuencia Conservada , Evolución Molecular , Genes Reporteros , Células HeLa , Humanos , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Unión Proteica , Ribonucleoproteínas/metabolismo , Factor de Empalme U2AF
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