A YAC library enriched for telomere clones was constructed and screened for the human telomere-specific repeat sequence (TTAGGG). Altogether 196 TYAC library clones were studied: 189 new TYAC clones were isolated, 149 STSs were developed for 132 different TYACs, and 39 P1 clones were identified using 19 STSs from 16 of the TYACs. A combination of mapping methods including fluorescence in situ hybridization, somatic cell hybrid panels, clamped homogeneous electric fields, meiotic linkage, and BLASTN sequence analysis was utilized to characterize the resource. Forty-five of the TYACs map to 31 specific telomere regions. Twenty-four linkage markers were developed and mapped within 14 proterminal regions (12 telomeres and 2 terminal bands). The polymorphic markers include 12 microsatellites for 10 telomeres (1q, 2p, 6q, 7q, 10p, 10q, 13q, 14q, 18p, 22q) and the terminal bands of 11q and 12p. Twelve RFLP markers were identified and meiotically mapped to the telomeres of 2q, 7q, 8p, and 14q. Chromosome-specific STSs for 27 telomeres were identified from the 196 TYACs. More than 30,000 nucleotides derived from the TYAC vector-insert junction regions or from regions flanking TYAC microsatellites were compared to reported sequences using BLASTN. In addition to identifying homology with previously reported telomere sequences and human repeat elements, gene sequences and a number of ESTs were found to be highly homologous to the TYAC sequences. These genes include human coagulation factor V (F5), Wee1 protein tyrosine kinase (WEE1), neurotropic protein tyrosine kinase type 2 (NTRK2), glutathione S-transferase (GST1), and β tubulin (TUBB). The TYAC/P1 resource, derivative STSs, and polymorphisms constitute an enabling resource to further studies of telomere structure and function and a means for physical and genetic map integration and closure.
Bibliographical noteFunding Information:
We thank Dr. Thomas Cremer in particular for guidance in FISH analyses, for advice over the course of the project, and for reviewing the manuscript prior to submission. We thank M. V. Olson for discussions and advice during the early phases of this work. We also thank M. V. Olson and H. Riethman for providing 7 TYAC clones, a number of TYAC cosmid subclones, and ligation mixtures from which the TYAC library was constructed. The technical assistance of Mary Pat Leicke from the Olson laboratory is gratefully acknowledged as are the contributions from the H.D-K. laboratory research staff, administrative staff, and students, in particular, Mary Akin, Stephanie Amen, Suzanne Cole, Meghan Dierks, William Dilley, Jim Dutchik, Michelle Lacy, Martin Lee, Jing Li, Brian Luketin, Bill Malone, Mark Schaller, Ed Shelton, and Chris Tierney. This work was supported in part by NIH Grants HG00100 and P41-HG01066 (to H.D-K.).