1995 Fiscal Year Final Research Report Summary
RNases associated with self-incompatibility in plants.
| Project/Area Number |
06044141
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Osaka University |
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Principal Investigator |
SAKIYAMA Fumio Institute for Protein Research, Osaka University, Professor, 蛋白質研究所, 教授 (40029947)
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| Co-Investigator(Kenkyū-buntansha) |
李 紹良 大阪大学, 蛋白質研究所, 助手 (40252720)
SATO Mamoru Institute for Protein Research, Osaka University Assistant Professor, 蛋白質研究所, 助手 (60170784)
NORIOKA Shigemi Institute for Protein Research, Osaka University Associate Professor, 蛋白質研究所, 助教授 (70198638)
CLARKE Adrienne e School of Botany, Melbourne University, Professor, 植物学部, 教授
LI Shao-liang Institute for Protein Research, Osaka University Assistant Professor
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| Project Period (FY) |
1994 – 1995
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| Keywords | Self-incompatibility / Ribonuclease / Japanese pear / Nicotiana alata / S-RNase / Primary structure / Tertiary structure |
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| Research Abstract |
Primary Structures of S-RNases of Japanese Pear - We purified four S-RNases (S2-, S3-, S4- and S5-RNases) from the styles of Japanese pear by a series of CM-cellulose and hydroxyapatite chromatographies. Subsequently, we determined the entire sequence of S4-RNase and the partial sequences of S3-RNase (95% of the entire sequence) and S5-RNase (95% of the entire sequence) by conventional protein sequencing. Since S2-RNase was isolated at much lower yield than those of the other S-RNases, only 25% of its entire sequence was determined by protein sequencing of peptides separated from lysylendopeptidase-digest of CM-S2-RNase. We then cloned cDNA encoding S2-RNase based on its partial sequence and deduced the entire amino acid sequence from its nucleotide sequence. By sequence comparison between these S-RNases and the solanaceous S-RNases, we found structure motifs common to all the S-RNases and stuctural features specific for the S-RNases of Japanese pear, and presumed regions specific for the S-allele which may be recognized by unidentified S-product in a pollen.
X-Ray Crystallographic Study of S1-RNase of Nicotiana alata - S1-RNase of Nicotiana alata was crystallized by hanging drop method using PEG-6000. The S1-RNase crystal was monoclinic, P2_1, a=80.3 , b=77.9, c=68.3 , beta=90.0ﾟ.This crystal was found to be available for tertiary structure analysis of S1-RNase.
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