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J. Biol. Chem., Vol. 277, Issue 39, 36380-36386, September 27, 2002
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From the Division of Applied Bioscience, Graduate School of
Agriculture, Hokkaido University, Sapporo 060-8589, Japan
Cystathionine
Identification of a Short Highly Conserved Amino
Acid Sequence as the Functional Region Required for
Posttranscriptional Autoregulation of the Cystathionine
-Synthase Gene in Arabidopsis*
,
,,
-synthase (CGS) catalyzes the
first committed step of Met biosynthesis in plants. We have previously
shown that expression of the gene for CGS is feedback-regulated at the level of mRNA stability, and that the amino acid sequence encoded by the first exon of the CGS gene itself is responsible for the regulation (Chiba, Y., Ishikawa, M., Kijima, F., Tyson, R. H., Kim, J., Yamamoto, A., Nambara, E., Leustek, T., Wallsgrove, R. M., and Naito, S. (1999) Science 286, 1371-1374). To
identify the functional region within CGS exon 1, deletion analysis was performed. The results showed that the 41-amino acid region of exon 1 highly conserved among plants is necessary and sufficient for the
regulation. Analyses of in vivo and in vitro
generated mutations that abolish the regulation identified the
functionally important amino acids as 11-13 residues within this
conserved region. The importance of these residues was confirmed by
deletion analysis within the conserved region. These studies identified the functional region of CGS exon 1 required for the
posttranscriptional autoregulation of the CGS gene as
(A)RRNCSNIGVAQ(I), with uncertainty of the first and last
residues. This sequence is almost perfectly conserved among CGS
sequences of higher plants but cannot be found elsewhere in the public databases.
*
This work was supported by Grants-in-Aid for Scientific
Research from the Ministry of Education, Culture, Sports, Science and
Technology of Japan 12138201 and 13440233 and by the "Research for
the Future" program from the Japanese Society for the Promotion of
Science (JSPS) Grant JSPS-RFTF97L00601.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.
Present address: Morinaga Research Institute, Tsurumi, Yokohama
230-8504, Japan.
§
Present address: Graduate School of Environmental Earth Science,
Hokkaido University, Sapporo 060-0810, Japan.
¶
JSPS Research Fellow.
Present address: Fukujuen CHA Research Center, Kyoto 619-0223, Japan.
**
Present address: Graduate School of Biological Sciences, Nara
Institute of Science and Technology, Ikoma 630-0101, Japan.
JSPS Research Associate for the "Research for the Future" program.
§§
Present address: Delaware Biotechnology Institute, University of
Delaware, 15 Innovation Way, Newark, DE 19711.
¶¶
To whom correspondence should be addressed. Fax:
81-11-706-4932; E-mail: naito@abs.agr.hokudai.ac.jp.
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