Telomere structure in Euplotes crassus: characterization of DNA-protein interactions and isolation of a telomere-binding protein.
The nucleoprotein structure of telomeres from Euplotes crassus was studied by using nuclease and chemical footprinting. The macronuclear telomeres were found to exist as DNA-protein complexes that are resistant to micrococcal nuclease digestion. Each complex encompassed 85 to 130 base pairs of macronuclear DNA and appeared to consist of two structural domains that are characterized by dissimilar DNA-protein interactions. Dimethyl sulfate footprinting demonstrated that very sequence-specific and salt-stable interactions occur in the most terminal region of each complex. DNase I footprinting indicated that DNA in the region 30 to 120 base-pairs from the 5' end lies on a protein surface; the interactions in this region of the complex are unlikely to be sequence specific. A 50-kilodalton telomere-binding protein was isolated. Binding of this protein protected telomeric DNA from BAL 31 digestion and gave rise to many of the sequence-specific DNA-protein interactions that were observed in vivo. The telomeric complexes from E. crassus were very similar in overall structure to the complexes found at Oxytricha telomeres. However, telomeric complexes from the two ciliates showed significant differences in internal organization. The telomeric DNA, the telomere-binding proteins, and the resultant DNA-protein interactions were all somewhat different. The telomere-binding proteins from the two ciliates were found to be less closely conserved than might have been expected. It appears that the proteins are tailored to match their cognate telomeric DNA.
Price
C M
CM
Department of Chemistry, University of Nebraska-Lincoln 68588-0304.
D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000302:isolation & purification*; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000648:ultrastructure; D002875:Chromosomes / Q000502:physiology* / Q000648:ultrastructure; D004247:DNA / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D003850:Deoxyribonuclease I; D056890:Eukaryota / Q000235:genetics / Q000502:physiology*; D008836:Micrococcal Nuclease; D008969:Molecular Sequence Data
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D004247:DNA; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease