Cloning and characterisation of the chicken gene encoding the telomeric protein TRF2

The telomere-associated protein TRF2 binds as a homodimer to double-stranded (TTAGGG)n arrays in vitro and localises to chromosome ends in vivo. Inhibition of TRF2 in human cell lines and recent electron microscopy analyses suggest that TRF2 plays a crucial role in the maintenance of telomere integr...

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Bibliographic Details
Published in:	Gene Vol. 239; no. 1; pp. 81 - 90
Main Authors:	Konrad, Jonathan P, Mills, Walter, Easty, David J, Farr, Christine J
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 18-10-1999
Subjects:	(TTAGGG)n Amino Acid Sequence Animals Base Sequence Cell Line, Transformed Chickens Chromosome Mapping Cloning, Molecular Codon, Initiator Conserved Sequence Dimerisation domain DNA, Complementary - chemistry DNA, Complementary - genetics DNA, Complementary - isolation & purification DNA-Binding Proteins - genetics Exons Gene Expression Regulation, Developmental Genes - genetics Humans Introns Mice Molecular Sequence Data myb-related DNA binding motif Protein Biosynthesis Sequence Alignment Sequence Analysis, DNA Sequence Homology, Amino Acid Telobox Telomere Telomere - genetics Telomeric Repeat Binding Protein 2 Tumor Cells, Cultured Vertebrate Vertebrates - genetics TRF2, gene encoding telomeric-repeat binding factor 2 ( Karlseder et al., 1999) cDNA, complementary DNA MRC, Medical Research Council of Gt Britain 3′, three prime end (TTAGGG)n (poly)A+, polyadenylated RNA C, cytidine A, adenosine Telomere ATM, ataxia telangiectasia CK, casein kinase cAMP, cyclic adenosine monophosphate DB, database dT, deoxythymidine G, guanosine NLS, nuclear localisation signal FISH, fluorescence in situ hybridisation SV, SV40 virus T, thymidine SDS, sodium dodecyl sulphate bps, base pairs mRNA, messenger RNA PKC, protein kinase C ORF, open reading frame SSC, 0.15 M NaCl/0.015 M Na3.citrate pH 7.6 myb-related DNA binding motif oligo, oligonucleotide GAPDH, glyceraldehyde phosphate dehydrogenase DNA nts, nucleotides Vertebrate Dimerisation domain 3′ UNT, untranslated transcript CMV, cytomegalovirus PCR, polymerase chain reaction RACE, rapid amplification of cDNA ends TRF1, gene encoding telomeric-repeat binding factor 1 ( Karlseder et al., 1999) Telobox t loop, denotes telomeric DNA forming large duplex loops ( Griffith et al., 1999)
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Summary:	The telomere-associated protein TRF2 binds as a homodimer to double-stranded (TTAGGG)n arrays in vitro and localises to chromosome ends in vivo. Inhibition of TRF2 in human cell lines and recent electron microscopy analyses suggest that TRF2 plays a crucial role in the maintenance of telomere integrity. To study the role of TRF2 in vertebrate telomere biology using an alternative model system, we report the isolation and characterisation of the chicken TRF2 locus. The TRF2 protein is highly conserved between mammals and birds, particularly within the dimerisation and myb-type DNA binding domains. However, the chicken ORF predicts an additional protein domain consisting of 15 copies of a degenerate 13 amino acid repeat. Indirect immunofluorescence reveals the localisation of a FLAG-tagged version of the chicken TRF2 protein at chromosome ends in both chicken and human cells suggesting that the protein is functionally conserved.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0378-1119 1879-0038
DOI:	10.1016/S0378-1119(99)00374-1