THE TELOMERE AND TELOMERASE - NUCLEIC ACID-PROTEIN COMPLEXES ACTING IN A TELOMERE HOMEOSTASIS SYSTEM - A REVIEW

The tandemly repeated DNA sequence of telomeres is typically specified by the ribonucleoprotein enzyme telomerase. Telomerase copies part of its intrinsic RNA moiety to synthesize one strand of the telomeric re peat DNA. Recent work, taken together with many observations over the past years, has led to the concept of a telomere homeostasis system. W e have analyzed the interplay between two key physical components of t his system: structural components of the telomere itself and of telome rase. Here we review some of these recent studies, The experimental me thod used in common in these studies was to make mutations in the temp late sequence of telomerase RNA, which caused various phenotypes. Firs t, mutating specific residues in the ciliate Tetrahymena thermophila a nd yeast showed that these residues are required for critical aspects of the enzymatic action of telomerase, Second, certain mutated telomer ic sequences caused a strong anaphase block in Tetrahymena micronuclei . Third, specific template mutations in the telomerase RNA gene led to varying degrees of telomers elongation in Tetrahymena and the yeast K luyveromyces lactis. For some of the K. lactis mutations, the loss of length unregulated elongation was directly related to loss of binding to K. lactis Rap1p protein. Using K. lactis carrying alterations in th e telomerase RNA template, and in the gene encoding the Rap1p protein, we found that a crucial determinant of telomere length homeostasis is the nature of the duplex DNA-Rap1p protein complex on the very end re peat of the telomere. We propose that this complex plays a key role in regulating access of telomerase to the telomere.