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Structure and Targeting of Precursor MicroRNA-31: From Mechanism to Application
Structure and Targeting of Precursor MicroRNA-31: From Mechanism to Application
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211152059
- ISBN
- 9798382739632
- DDC
- 574
- 저자명
- Ma, Sicong.
- 서명/저자
- Structure and Targeting of Precursor MicroRNA-31: From Mechanism to Application
- 발행사항
- [Sl] : University of Michigan, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 264 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-12, Section: B.
- 주기사항
- Advisor: Keane, Sarah.
- 학위논문주기
- Thesis (Ph.D.)--University of Michigan, 2024.
- 초록/해제
- 요약As an essential post-transcriptional regulator of gene expression, microRNA (miRNA) levels must be strictly maintained. The biogenesis of many miRNAs is regulated by trans-acting protein partners, which exert control through a variety of mechanisms, including remodeling of the RNA structure and recruitment of RNA processing or modifying enzymes. MicroRNA-31 (miR-31) functions as an oncogene in numerous cancers, and interestingly, its biogenesis is not known to be regulated by protein factors. Therefore, I investigated if the intrinsic structural and dynamical properties of the miR-31 precursor element, pre-miR-31, can provide a mechanism by which its biogenesis is regulated. Base pair mismatches are a common feature of primary and precursor miRNAs. In this thesis, I characterized the base pair mismatches within pre-miR-31 and found the C•A mismatch within the stem of pre-miR-31 to be strongly pH sensitive and, stabilizing the RNA structure at near physiological pH. Next, I investigated the role of distinct structural elements within pre-miR-31 in regulating processing by the Dicer/TRBP complex. I found that both the apical loop size and structure at the Dicing site are key elements for discrimination by the Dicer/TRBP complex. Interestingly, our NMR-derived structure revealed the presence of a triplet of base pairs, or junction region, that link the Dicer cleavage site and the apical loop. My mutational analysis in this region revealed that the stability of the junction region strongly influenced processing by the Dicer/TRBP complex. Based on these findings, I developed a new type of antisense oligonucleotide (ASO) that specifically targets the junction region of pre-miR-31 to inhibit Dicer/TRBP processing. Furthermore, I demonstrated that this new type of ASO design is broadly applicable to other junction containing pre-miRNAs, which account for ~20 % of human pre-miRNAs, and function to reduce the Dicer/TRBP cleavage of this family of pre-miRNAs. These studies enhance our understanding of RNA structure based ASO design and development. The results in this thesis enrich our understanding of the active role that RNA structure plays in regulating miRNA biogenesis, which has direct implications for the control of gene expression. This study further points out that RNA structure is not a passive element in the protein enzymatic steps. Rather, the RNA structural elements play important roles in regulating processing by Dicer/TRBP. Furthermore, my thesis work provides a new approach for antisense oligonucleotide design by targeting microRNA biogenesis at a step upstream of the traditional anti-microRNA antisense oligonucleotide design strategy. This new type of antisense oligonucleotide allows intervention at an early stage of miRNA biogenesis and may lead to a novel treatment by selectively inhibiting disease-related pre-miRNAs.
- 일반주제명
- Biochemistry
- 일반주제명
- Biophysics
- 일반주제명
- Molecular biology
- 키워드
- MicroRNA-31
- 키워드
- RNA
- 키워드
- Biogenesis
- 기타저자
- University of Michigan Biophysics
- 기본자료저록
- Dissertations Abstracts International. 85-12B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520250211152059
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■020 ▼a9798382739632
■035 ▼a(MiAaPQ)AAI31349009
■035 ▼a(MiAaPQ)umichrackham005515
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aMa, Sicong.
■24510▼aStructure and Targeting of Precursor MicroRNA-31: From Mechanism to Application
■260 ▼a[Sl]▼bUniversity of Michigan▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a264 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-12, Section: B.
■500 ▼aAdvisor: Keane, Sarah.
■5021 ▼aThesis (Ph.D.)--University of Michigan, 2024.
■520 ▼aAs an essential post-transcriptional regulator of gene expression, microRNA (miRNA) levels must be strictly maintained. The biogenesis of many miRNAs is regulated by trans-acting protein partners, which exert control through a variety of mechanisms, including remodeling of the RNA structure and recruitment of RNA processing or modifying enzymes. MicroRNA-31 (miR-31) functions as an oncogene in numerous cancers, and interestingly, its biogenesis is not known to be regulated by protein factors. Therefore, I investigated if the intrinsic structural and dynamical properties of the miR-31 precursor element, pre-miR-31, can provide a mechanism by which its biogenesis is regulated. Base pair mismatches are a common feature of primary and precursor miRNAs. In this thesis, I characterized the base pair mismatches within pre-miR-31 and found the C•A mismatch within the stem of pre-miR-31 to be strongly pH sensitive and, stabilizing the RNA structure at near physiological pH. Next, I investigated the role of distinct structural elements within pre-miR-31 in regulating processing by the Dicer/TRBP complex. I found that both the apical loop size and structure at the Dicing site are key elements for discrimination by the Dicer/TRBP complex. Interestingly, our NMR-derived structure revealed the presence of a triplet of base pairs, or junction region, that link the Dicer cleavage site and the apical loop. My mutational analysis in this region revealed that the stability of the junction region strongly influenced processing by the Dicer/TRBP complex. Based on these findings, I developed a new type of antisense oligonucleotide (ASO) that specifically targets the junction region of pre-miR-31 to inhibit Dicer/TRBP processing. Furthermore, I demonstrated that this new type of ASO design is broadly applicable to other junction containing pre-miRNAs, which account for ~20 % of human pre-miRNAs, and function to reduce the Dicer/TRBP cleavage of this family of pre-miRNAs. These studies enhance our understanding of RNA structure based ASO design and development. The results in this thesis enrich our understanding of the active role that RNA structure plays in regulating miRNA biogenesis, which has direct implications for the control of gene expression. This study further points out that RNA structure is not a passive element in the protein enzymatic steps. Rather, the RNA structural elements play important roles in regulating processing by Dicer/TRBP. Furthermore, my thesis work provides a new approach for antisense oligonucleotide design by targeting microRNA biogenesis at a step upstream of the traditional anti-microRNA antisense oligonucleotide design strategy. This new type of antisense oligonucleotide allows intervention at an early stage of miRNA biogenesis and may lead to a novel treatment by selectively inhibiting disease-related pre-miRNAs.
■590 ▼aSchool code: 0127.
■650 4▼aBiochemistry
■650 4▼aBiophysics
■650 4▼aMolecular biology
■653 ▼aMicroRNA-31
■653 ▼aRNA
■653 ▼aAntisense oligonucleotide
■653 ▼aBiogenesis
■690 ▼a0786
■690 ▼a0487
■690 ▼a0307
■71020▼aUniversity of Michigan▼bBiophysics.
■7730 ▼tDissertations Abstracts International▼g85-12B.
■790 ▼a0127
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17162825▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
