The Euplotes La motif protein p43 has properties of a telomerase-specific subunit.
Telomerase is a specialized reverse transcriptase synthesizing DNA repeats at telomeres. In addition to the RNA and catalytic protein components, telomerase from the ciliate Euplotes aediculatus contains the subunit p43. This protein is homologous to the La autoantigen, functioning in maturation of RNA polymerase III transcripts. Here we provide evidence that p43 is primarily associated with the telomerase ribonucleoprotein in vivo. Recombinant p43 binds telomerase RNA with low-nanomolar affinity in vitro, recognizing stem I and adjacent nucleotides or structures in the core of the RNA. Unlike authentic La proteins, p43 does not bind strongly to RNA polymerase III precursor transcripts and does not exhibit a marked binding preference for 3'-terminal oligouridylate residues. In isolated macronuclei, p43 largely colocalizes with telomerase RNA in discrete foci. These findings suggest that p43 is not the Euplotes La protein but instead plays a dedicated role in telomerase assembly and/or function. Thus, p43 joins the telomerase reverse transcriptase and the yeast proteins Est1p and Est3p as the only telomerase-specific proteins identified so far.
Aigner
Stefan
S
Department of Chemistry and Biochemistry and Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 80309-0215, USA.
Postberg
Jan
J
Lipps
Hans J
HJ
Cech
Thomas R
TR
D000818:Animals; D001324:Autoantigens; D001483:Base Sequence; D001665:Binding Sites; D016798:Ciliophora / Q000235:genetics / Q000378:metabolism; D016054:DNA, Protozoan / Q000235:genetics; D016812:Euplotes / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D021122:Protein Subunits; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019098:Telomerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013768:Tetrahymena / Q000235:genetics / Q000378:metabolism; D000097585:SS-B Antigen
D001324:Autoantigens; D016054:DNA, Protozoan; D021122:Protein Subunits; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D019098:Telomerase; D000097585:SS-B Antigen