Title

Characterization of cohesin dynamics mediated by STAG variants and SMC3 acetylation

Alternative Title

STAG 변이체와 SMC3 아세틸화에 의한 코헤신 동역학의 특성 비교

DGIST Authors

Minseok Seo ; Yoori Kim ; Jin Hae Kim

Advisor

김유리

Co-Advisor(s)

Jin Hae Kim

Issued Date

2026

Awarded Date

2026-08-01

Type

Thesis

Description

cohesin, STAG1/2, SMC3 acetylation, insect cell-based purification, cohesin dynamics

Table Of Contents

Ⅰ. Introduction 1
II. Materials and Methods 11
2.1 Expression plasmids for cohesin subunits 11
2.2 Site-directed mutagenesis for SMC3 K105Q / K106Q （SMC3 QQ mutant） 11
2.3 Generation of STAG2 expression plasmid 12
2.4 Blue-white screening 13
2.5 Isolation of recombinant Bacmid DNA and analyze by PCR 14
2.6 Baculovirus production and amplification in Sf9 insect cells 15
2.7 Protein expression in High-Five insect cells 16
2.8 Cohesin variants complex purification 16
2.9 Electrophoretic mobility shift assay （EMSA） 18
2.10 ATPase assay 19
Ⅲ. Results 21
3.1 Cohesin complex expression using Bac-to-Bac baculovirus system 21
3.2 Purification of cohesin variant complexes 27
3.3 SMC3 acetylation-mimic state reduces cohesin ATPase activity 32
3.4 STAG variants and SMC3 acetylation regulate cohesin DNA binding 34
3.5 STAG1 and STAG2 directly bind DNA with distinct binding behavior 36
3.6 DNA binding preferences of STAG1 and STAG2 39
Ⅳ. Discussion 41
References 47
Summary in Korean 57

URI

https://scholar.dgist.ac.kr/handle/20.500.11750/60776
http://dgist.dcollection.net/common/orgView/200001018065

DOI

10.22677/THESIS.200001018065

Degree

Master

Department

Department of New Biology

Publisher

DGIST

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