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Regulation of RNA processing in B lymphocyte differentiation

Research Project

Project/Area Number 07044249
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SHIMIZU Akira  Center for Molecular Biology and Genetics, Kyoto University, 遺伝子実験施設, 教授 (00162694)

Co-Investigator(Kenkyū-buntansha) STORB Ursula Beate  Department of Molecular Genetics and Cell Biology, The University of Chicago, 分子遺伝学・細胞生物学科, 教授
MARTIN Teren  シカゴ大学, 分子遺伝学・細胞生物学科, 教授
AGATA Yasutoshi  Center for Molecular Biology and Genetics, Kyoto University, 遺伝子実験施設, 助手 (60263141)
MARTIN Terence Edwin  Department of Molecular Genetics and Cell Biology, The University of Chicago
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsIg class Switching / Ig gene / S region / Gene rearrangement / germ-line transcript / trans-splicing / trans-mRNA / Multiple isotype expression / アレルギー / CD40リガンド
Research Abstract

We cloned the constant (C) region gene of chicken immunoglobulin (Ig)mu chain. Nucleotide sequence analysis revealed that there are two types of tandem repetitive sequences apanning up to a few kilobases locating just upstream of the first exon of C_<mu> gene. Both of the repeats consist of unit sequences of 5 base long, resembling to the S_<mu> region of mammalian gene. One of the two types, closer to the C_<mu> gene consists of tandem repeats of (C/T) (C/A) CAG complementary to the mammalian S_<mu> region. These findings suggest the DNA recombination during the class switching in chicken, and importance of the repetitive nature of S regions.
We next tested whether the Ig trans-mRNAs, which consist of a variable (V) region of a transgene and the C regions of endogenous isotypes, can be expressed from a transgene lacking S region which is necessary for the DNA recombination of class switching. Spleen cells of C line mouse, a transgenic mouse carrying a rearranged mouse gamma2b gene from which the S_<mu> and S_<gamma2b> regions were completely deleted during the construction the transgene, were stimulated with LPS and IL-4 to induce the trans-mRNA expression. Upon the stimulation, specific induction of gamma1 and epsilon trans-mRNAs were detected by RT-PCR using specific primers for transgenic V region and endogenous C region genes. Structures of the detected PCR products were confirmed by nested PCRs with specific, inner primers and their direct sequencing using an automated sequencer. Our data indicate that the trans-mRNA expression is independent of S region mediated mechanism, i.e., DNA (S-S) recombination. This also supports our previous hypothesis that Ig trans-mRNAs are expressed by RNA mediated mechanism, i.e., trans-splicing, and thus multiple isotypes of Ig can be expressed through this mechanism.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Kitao,H.,Arakawa,H.,Yamagishi,H.and Shimizu,A.: "Chiken immunoglobulin μ-chain gene:germline organization and tandem repeats characteristic of class switch recombination." Immunology Letters. 52. 99-104 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 清水章: "抗体クラススイッチの分子機構とIgE" 日本臨床. 54. 440-445 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 清水章: "抗体機能の発現とその制御" 蛋白質 核酸 酸素. 40. 2107-2117 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitao, H., Arakawa, H., Yamagishi, H.and Simizu, A.: "Chicken immunoglobulin mu-chain gene : germline organization and tandem repeats characteristic of class switch recombination." Immunology Letters. 52. 99-104 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, A.: "Molecular mechanisms for immunoglobulin class switching and IgE production." Nippon-Rinsho. 54. 440-445 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, A.: "Analysis of regulatory mechanism for humoral immunity using transgenic mice of rearranged immunoglobulin genes." Tanpakushitsu-Kakusan-Koso. 40. 2107-2117 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitao,H.,Arakawa,H.,Yamagishi,H.and Shimizu,A.: "Chicken immunoglobulin μ-chain gene:germline organization and tandem repeats characteristics of class switch recombination" Immunolgy Letters. 52. 99-104 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 清水章: "抗体機能の発現とその制御" 蛋白質 核酸 酵素. 40. 2107-2117 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 清水章: "抗体クラススイッチの分子機構とIgE" 日本臨床. 54. 440-445 (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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