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J. Biol. Chem., Vol. 277, Issue 9, 6903-6914, March 1, 2002
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From the Dipartimento di Biochimica e Biologia Molecolare,
Università di Parma, I-43100 Parma, Italy
The SCR1 gene, coding for the 7SL RNA
of the signal recognition particle, is the last known class III gene of
Saccharomyces cerevisiae that remains to be characterized
with respect to its mode of transcription and promoter organization. We
show here that SCR1 represents a unique case of a non-tRNA
class III gene in which intragenic promoter elements (the
TFIIIC-binding A- and B-blocks), corresponding to the D and T
Intragenic Promoter Adaptation and Facilitated RNA Polymerase III
Recycling in the Transcription of SCR1, the 7SL RNA Gene of
Saccharomyces cerevisiae*
,
C arms
of mature tRNAs, have been adapted to a structurally different small
RNA without losing their transcriptional function. In fact, despite the
presence of an upstream canonical TATA box, SCR1
transcription strictly depends on the presence of functional, albeit
quite unusual, A- and B-blocks and requires all the basal
components of the RNA polymerase III transcription
apparatus, including TFIIIC. Accordingly, TFIIIC was found to protect
from DNase I digestion an 80-bp region comprising the A- and B-blocks.
B-block inactivation completely compromised TFIIIC binding and
transcription capacity in vitro and in vivo. An
inactivating mutation in the A-block selectively affected TFIIIC
binding to this promoter element but resulted in much more dramatic
impairment of in vivo than in vitro
transcription. Transcriptional competition and nucleosome disruption
experiments showed that this stronger in vivo defect is due
to a reduced ability of A-block-mutated SCR1 to compete
with other genes for TFIIIC binding and to counteract the assembly of
repressive chromatin structures through TFIIIC recruitment. A kinetic
analysis further revealed that facilitated RNA polymerase III
recycling, far from being restricted to typical small sized class III
templates, also takes place on the 522-bp-long SCR1 gene,
the longest known class III transcriptional unit.
*
This work was supported by grants from the Ministry of
Education, University, and Research (Rome, Italy; Cofin-PRIN
program).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence may be addressed. Tel.: 39-0521-905646;
Fax: 39-0521-905151; E-mail: gdieci@unipr.it.
§
To whom correspondence may be addressed. Tel.: 39-0521-905646; Fax:
39-0521-905151; E-mail: s.ottonello@unipr.it.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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