DNA bound by the Oxytricha telomere protein is accessible to telomerase and other DNA polymerases.
Macronuclear telomeres in Oxytricha exist as DNA-protein complexes in which the termini of the G-rich strands are bound by a 97-kDa telomere protein. During telomeric DNA replication, the replication machinery must have access to the G-rich strand. However, given the stability of telomere protein binding, it has been unclear how this is accomplished. In this study we investigated the ability of several different DNA polymerases to access telomeric DNA in Oxytricha telomere protein-DNA complexes. Although DNA bound by the telomere protein is not degraded by micrococcal nuclease or labeled by terminal deoxynucleotidyltransferase, this DNA serves as an efficient primer for the addition of telomeric repeats by telomerase, a specialized RNA-dependent DNA polymerase (ribonucleoprotein reverse transcriptase), EC 2.7.7.49. Moreover, in the presence of a suitable complementary C-rich DNA template, AMV reverse transcriptase and the E. coli Klenow fragment will also elongate DNA bound by the telomere protein. These findings indicate that the 3' terminus and the Watson-Crick base pairing positions are exposed in the protein complex. We propose that the telomere protein can serve a dual role at the telomere by protecting the DNA phosphate backbone from degradation while simultaneously exposing the DNA bases for replication.
Shippen
D E
DE
Department of Microbiology and Immunology, University of California, San Francisco 94143.
Blackburn
E H
EH
Price
C M
CM
D000818:Animals; D001483:Base Sequence; D002843:Chromatin / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D004256:DNA Polymerase I / Q000378:metabolism*; D016054:DNA, Protozoan / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016813:Oxytricha; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D016615:Telomere / Q000378:metabolism*
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D016054:DNA, Protozoan; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D015800:Protozoan Proteins; D004253:DNA Nucleotidylexotransferase; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I