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Molecular and Cellular Biology, July 2002, p. 5248-5256, Vol. 22, No. 14
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.14.5248-5256.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Lsm Proteins Are Required for Normal Processing of Pre-tRNAs and Their Efficient Association with La-Homologous Protein Lhp1p

Joanna Kufel, Christine Allmang,,{dagger} Loredana Verdone,,{ddagger} Jean D. Beggs, and David Tollervey*

Wellcome Trust Centre for Cell Biology, The University of Edinburgh, Edinburgh EH9 3JR, United Kingdom

Received 11 February 2002/ Returned for modification 20 March 2002/ Accepted 18 April 2002

Depletion of any of the five essential proteins Lsm2p to Lsm5p and Lsm8p leads to strong accumulation of all tested unspliced pre-tRNA species, as well as accumulation of 5' and 3' unprocessed species. Aberrant 3'-extended pre-tRNAs were detected, presumably due to stabilization of transcripts that fail to undergo correct transcription termination, and the accumulation of truncated tRNA fragments was also observed. Tandem affinity purification-tagged Lsm3p was associated with pre-tRNA primary transcripts and, less efficiently, with other unspliced pre-tRNA intermediates but not mature tRNAs. Association of the Saccharomyces cerevisiae La homologue Lhp1p with pre-tRNAs was reduced approximately threefold on depletion of Lsm3p or Lsm5p. The association of Lhp1p with larger RNA polymerase III transcripts, pre-RNase P RNA and the signal recognition particle RNA (scR1), was more drastically reduced. The impaired pre-tRNA processing seen on Lsm depletion is not, however, due solely to reduced Lhp1p association as evidenced by analysis of lhp1-{Delta} strains depleted of Lsm3p or Lsm5p. These data are consistent with roles for an Lsm complex as a chaperone that facilitates the efficient association of pre-tRNA processing factors with their substrates.


* Corresponding author. Mailing address: Wellcome Trust Centre for Cell Biology, King's Buildings, The University of Edinburgh, Edinburgh EH9 3JR, United Kingdom. Phone: 44 131 650 7092. Fax: 44 131 650 7040. E-mail: d.tollervey{at}ed.ac.uk.

{dagger} Present address: Institut de Biologie Moleculaire et Cellulaire, UPR 9002 du CNRS, 67084 Strasbourg Cedex, France.

{ddagger} Present address: Department of Molecular Biology, University of Rome "La Sapienza," 00185 Rome, Italy.


Molecular and Cellular Biology, July 2002, p. 5248-5256, Vol. 22, No. 14
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.14.5248-5256.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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