N-terminal domain of yeast telomerase reverse transcriptase: recruitment of Est3p to the telomerase complex.
Telomerase is a reverse transcriptase that maintains chromosome ends. The N-terminal half of the catalytic protein subunit (TERT) contains three functional domains (I, II, and III) that are conserved among TERTs but not found in other reverse transcriptases. Guided by an amino acid sequence alignment of nine TERT proteins, mutations were introduced into yeast TERT (Est2p). In support of the proposed alignment, mutation of virtually all conserved residues resulted in loss-of-function or temperature sensitivity, accompanied by telomere shortening. Overexpression of telomerase component Est3p led to allele-specific suppression of the temperature-sensitive mutations in region I, suggesting that Est3p interacts with this protein domain. As predicted by the genetic results, a lethal mutation in region I resulted in loss of Est3p from the telomerase complex. We conclude that Est2p region I is required for the recruitment of Est3p to yeast telomerase. Given the phylogenetic conservation of region I of TERT, this protein domain may provide the equivalent function in all telomerases.
Friedman
Katherine L
KL
Vanderbilt University, Department of Biological Sciences, Nashville, Tennessee 37235, USA. katherine.friedman@vanderbilt.edu
Heit
Jeremy J
JJ
Long
David M
DM
Cech
Thomas R
TR
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D006358:Hot Temperature; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D019098:Telomerase / Q000378:metabolism*; D015003:Yeasts / Q000201:enzymology* / Q000378:metabolism
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; C099024:EST1 protein, S cerevisiae; C491580:EST3 protein, S cerevisiae; D019098:Telomerase