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Mol Cell Biol, June 1998, p. 3201-3211, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
5' Processing of tRNA Precursors Can Be
Modulated by the Human La Antigen Phosphoprotein
Hao
Fan,1
John L.
Goodier,1
Joel R.
Chamberlain,2
David R.
Engelke,2,3 and
Richard J.
Maraia1,*
Laboratory of Molecular Growth Regulation,
National Institute of Child Health and Human Development, National
Institutes of Health, Bethesda, Maryland
20892-2753,1 and
Cellular and Molecular
Biology Program2 and
Department of
Biological Chemistry,3 University of Michigan,
Ann Arbor, Michigan 48109-0606
Received 24 November 1997/Returned for modification 18 January
1998/Accepted 6 March 1998
Eukaryotic precursor (pre)-tRNAs are processed at both ends prior
to maturation. Pre-tRNAs and other nascent transcripts synthesized by
RNA polymerase III are bound at their 3' ends at the sequence motif
UUUOH [3' oligo(U)] by the La antigen, a conserved
phosphoprotein whose role in RNA processing has been associated
previously with 3'-end maturation only. We show that in addition to its
role in tRNA 3'-end maturation, human La protein can also modulate 5' processing of pre-tRNAs. Both the La antigen's N-terminal RNA-binding domain and its C-terminal basic region are required for attenuation of
pre-tRNA 5' processing. RNA binding and nuclease protection assays with
a variety of pre-tRNA substrates and mutant La proteins indicate that
5' protection is a highly selective activity of La. This activity is
dependent on 3' oligo(U) in the pre-tRNA for interaction with the
N-terminal RNA binding domain of La and interaction of the C-terminal
basic region of La with the 5' triphosphate end of nascent pre-tRNA.
Phosphorylation of La is known to occur on serine 366, adjacent to the
C-terminal basic region. We show that this modification interferes with
the La antigen's ability to protect pre-tRNAiMet from
5' processing either by HeLa extract or purified RNase P but that it
does not affect interaction with the 3' end of pre-tRNA. These findings
provide the first evidence to indicate that tRNA 5'-end maturation may
be regulated in eukaryotes. Implications of triphosphate recognition is
discussed as is a role for La phosphoprotein in controlling
transcriptional and posttranscriptional events in the biogenesis of
polymerase III transcripts.
*
Corresponding author. Mailing address: Building 6, Room
416, LMGR, NICHD, NIH, 9000 Rockville Pike, Bethesda, MD 20892-2753. Phone: (301) 402-3567. Fax: (301) 480-6863. E-mail:
maraia{at}ncbi.nlm.nih.gov.

R.J.M. dedicates this research paper to the memory of Christopher
P. Cully.
Mol Cell Biol, June 1998, p. 3201-3211, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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