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Pin1 Interacts with Hepatitis B Virus Core Particle and Enhances HBV Replication

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dc.contributor.author권, 현중-
dc.date.accessioned2023-11-16T05:43:56Z-
dc.date.available2023-11-16T05:43:56Z-
dc.date.issued2023-
dc.identifier.urihttp://repository.ajou.ac.kr/handle/201003/26843-
dc.language.isoen-
dc.titlePin1 Interacts with Hepatitis B Virus Core Particle and Enhances HBV Replication-
dc.title.alternativeB형 간염바이러스 Core Particle과 Pin1의 상호 작용에 의한 HBV 증식 향상-
dc.typeThesis-
dc.identifier.urlhttp://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000033045-
dc.subject.keywordHepatitis B virus-
dc.subject.keywordHBV replication-
dc.subject.keywordCore particle-
dc.subject.keywordPPIase Pin1-
dc.subject.keywordPin1-Core particle interaction-
dc.subject.keywordB형 간염 바이러스-
dc.subject.keywordHBV 증식-
dc.subject.keyword코어 입자-
dc.subject.keywordPin1-
dc.subject.keywordPin1-코어입자 상호작용-
dc.description.degreeDoctor-
dc.contributor.department대학원 의생명과학과-
dc.contributor.affiliatedAuthor권, 현중-
dc.date.awarded2023-
dc.type.localTheses-
dc.citation.date2023-
dc.embargo.liftdate9999-12-31-
dc.embargo.terms9999-12-31-
dc.description.tableOfContentsI. INTRODUCTION 1

II. MATERIALS AND METHODS 5

A. Plasmids Used in the Study 5

B. Cell culture and Transfection 7

C. Establishment of Stable Cell Lines 8

D. Immunoprecipitation, SDS-PAGE, and Immunoblotting 8

E. Native Agarose Gel Electrophoresis and Immunoblotting 9

F. Analysis of Extracellular HBV Particles 10

G. HBV Infection 11

H. Investigation of Pin1-Core Particle Interaction 11

I. In Vitro Analysis of Pin1-Core Particle Interaction and Phosphorylation Dependency 12

J. Stability Analysis of the Core Particle 12

K. Southern, Northern, and In Situ Nucleic Acid Blotting 12

L. Extraction of cccDNA 13

III. RESULTS 16

A. The HBV Core Particle Is a Novel Binding Substrate of Pin1 16

B. Dynamic On-off Interaction Between Pin1 and the HBV Core Particle 25

C. Pin1 Binds Both to the Outside and Inside of the HBV Core Particle 26

D. The HBc S/TP Motifs Are Highly Conserved Among Ten Genotypes of Human HBV 29

E. The HBc 162TP, 164SP, and 172SP Motifs Are Important for the Pin1/Core Particle Interaction 34

F. HBc NTD S/TP Motifs Contribute to Core Particle Stability and Assembly 38

G. The 16Ser and 34Trp Substrate Binding Residues of Pin1 Increase Core Particle Stability Through the Pin1/Core Particle Interaction 42

H. Inhibition of Parvulin or PIN1 KD Downregulates HBV Replication 47

I. Pin1 Overexpression Upregulates HBV Replication 52

J. Fewer Pin1 Proteins Bind to HBV WT Core Particles Than to RT-defective or Priming-deficient Core Particles 58

K. Pin1 Overexpression Increases Secretion of Virions, whereas PIN1 KD Decreases Secretion of Virions 61

IV. Discussion 68

References 71
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