title: The telomerase-associated protein p43 is involved in anchoring telomerase in the nucleus.
abstract: Telomere replication of eukaryotic chromosomes is achieved by a specialized enzyme, the telomerase. Although the biochemistry of end-replication is well understood, little is known about the organization of the end-replication machinery, its regulation throughout the cell cycle or the biological function of the telomerase-associated proteins. Here we investigate the function of the telomerase-associated protein p43 within the macronucleus of the ciliated protozoa Euplotes. It has been shown that p43 binds in vitro to the RNA subunit of telomerase and shares homology with the La autoantigen family. It therefore has been suggested that it is involved in the assembly and/or nuclear retention of telomerase. We show that the p43-telomerase complex is bound to a subnuclear structure in vivo and is resistant to electroelution. Upon inhibition of p43 or telomerase expression by RNAi, which in this study was used for the first time in spirotrichs, this complex is no longer retained in the nucleus. Further analysis revealed that the p43-telomerase complex is bound to the nuclear matrix in vivo and that after inhibition of p43 expression, telomerase is released from this structure, strongly suggesting that p43 is involved in anchoring of telomerase in the nucleus. This is the first in vivo demonstration of the biological function of this telomerase-associated component involved in telomere replication and allows us to propose a model for the organization of the end-replication machinery in the eukaryotic cell.
authors.affiliation: Institute of Cell Biology, University Witten/Herdecke, Stockumer Strasse 10, D-58453 Witten, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Möllenbeck
authors.forename: Jan
authors.initials: J
authors.lastname: Postberg
authors.forename: Katrin
authors.initials: K
authors.lastname: Paeschke
authors.forename: Michael
authors.initials: M
authors.lastname: Rossbach
authors.forename: Franziska
authors.initials: F
authors.lastname: Jönsson
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Lipps
mesh_terms: D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D016054:DNA, Protozoan / Q000235:genetics; D016812:Euplotes / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances; D008954:Models, Biological; D011485:Protein Binding; D021122:Protein Subunits; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D019098:Telomerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D016054:DNA, Protozoan; D046911:Macromolecular Substances; D021122:Protein Subunits; D015800:Protozoan Proteins; D019098:Telomerase