| Project/Area Number |
63480416
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | The Nippon Dental University, Niigata |
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Principal Investigator |
SANADA Kazuo The Nippon Dental University, Niigata Department of Oral Biochemistry, Professor, 新潟歯学部, 教授 (70060423)
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| Co-Investigator(Kenkyū-buntansha) |
TERADA Akane The Nippon Dental University, Niigata Department of Oral Biochemistry, Research, 新潟歯学部, 助手 (60180080)
SHIMOMURA Hiromi The Nippon Dental University, Niigata Department of Oral Biochemistry, Assistant, 新潟歯学部, 助教授 (40139259)
SAITOH Eiichi The Nippon Dental University, Niigata Department of Oral Biochemistry, Lecturer, 新潟歯学部, 講師 (40120662)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | Saliva / Cysteine-proteinase inhibitor / Cystatin superfamily / Cystatin gene family / Gene expression / Nucleotide sequence / Human chromosome 20 / Molecular cloning / クロ-ニング / システインプロフア-ゼインヒビタ- / シスタチンC / シスタチンS / シスタチン遺伝子ファミリー / RFLPs / S型シスタチン / アミロイドーシス / 染色体DNA |
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| Research Abstract |
The superfamily of cysteine-proteinase inhibitors comprises structurally homologous proteins which are divided into at least three families: family I(the stefins),family II(the cystatins) and family II(the kininogens). Two classes of family II, salivary cystatins(cystatins S,SA and SN) and cystatin C differ in their irumunochemical and physicochemical properties, distribution and concentration. Physiological roles of these inhibitors could be the regulation of cysteine-proteinases of both host and bacterial origin. We have shown that the salivary-type cystatins are determined by the human cystatin gene family consist of at least 7 members, and isolated three cystatin genes, CST1 for cystatin SN, CST2 for cystatin SA and CSTP1 for a cystatin pseudogene.
To identify and characterize members of this gene family other than above three, we have screened the phage library containing HindIII digests of human genomic DNA using three DNA probes each including exon1,exon2 and exon3 of the CST1 ge
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ne. Four new members named CST2B(6.8kb),CST3(8.8kb),CST4(5.4kb) and CST5 (12.5kb) were isolated from this library. Nucleotide sequence analyses of these clones indicated that CST3 and CST4 code for the precursor forms of cystatin C and cystatin S,respectively. CST2B was considered to be an allele at the CST2 locus. Mapping and partial nucleotide sequence analysis suggested that CST5 does not code for any cystatin characterized so far.
In order to investigate the possible functions and tissue-specific regulations of cystatins,we examined the distribution of their RNAs in tissues of humans and monkeys. Several RNAs(O.8-1.3kb) were detested in human parotid gland,submandibular gland and bronchus, and lacrimal gland from monkeys. Thus it appears that the genes from the cystatin gene family are differentially expressed in several tissues.
Southern analysis of EcoRI digested DNAs from independent somatic cell hybrid clones hybridized to a probe from the CST1 gene demonstrated that all members of the cystatin gene family segregate with human chromosome 20.
In conclusion, two classes of family II cystatins, cystatin C and salivary- type cystatins are encoded by the different members of a multigene family which is localized on human chromosome 20. And also, inhibitors belonging to family II cystatin are produced by tissue-specific expression of the genes from the cystatin gene family. Less
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