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J. Biol. Chem., Vol. 276, Issue 1, 147-152, January 5, 2001
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From the Post-transcriptional RNA processing is an
important step in the regulation of chloroplast gene expression, and a
number of chloroplast ribonucleoproteins (cpRNPs) are likely to
be involved in this process. The major tobacco cpRNPs are composed of
five species: cp28, cp29A, cp29B, cp31, and cp33 and these are divided into three groups (I, II, and III). By immunoprecipitation, gel filtration, and Western blot analysis, we demonstrated that these cpRNPs are abundant stromal proteins that exist as complexes with ribosome-free mRNAs. Many ribosome-free psbA mRNAs
coprecipitate with cpRNPs, indicating that the majority of stromal
psbA mRNAs are associated with cpRNPs. In addition, an
in vitro mRNA degradation assay indicated that
exogenous psbA mRNA is more rapidly degraded in
cpRNP-depleted extracts than in nondepleted extracts. When the depleted
extract was reconstituted with recombinant cpRNPs, the psbA
mRNA in the extract was protected from degradation to a similar
extent as the psbA mRNA in the nondepleted extract. Moreover, restoration of the stabilizing activity varied following addition of individual group-specific cpRNPs alone or in combination. When the five cpRNPs were supplemented in the depleted extract, full
activity was restored. We propose that these cpRNPs act as stabilizing
factors for nonribosome-bound mRNAs in the stroma.
Chloroplast Ribonucleoproteins Function as a Stabilizing Factor
of Ribosome-free mRNAs in the Stroma*
,§,, and¶
Center for Gene Research, and the
¶ Graduate School of Human Informatics, Nagoya University,
Nagoya 464-8601, Japan
*
This work was supported by Grant-in-aid for Scientific
Research in Priority Areas No. 0927103 (to M. S.) from the Ministry of
Education, Science, Sports, and Culture of Japan.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 should be addressed. Tel/Fax: 81 52 789 4779; E-mail: sugita@info.human.nagoya-u.ac.jp.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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