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Nanyang Technological University

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion

1.3K Views

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08:24 min

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July 27th, 2021

DOI :

10.3791/61689-v

July 27th, 2021

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Jing Zhang*¹, Jing Huang*², Ishani Majumdar¹, Ryan C. James³, Julia Gichimu¹, Manikandan Paramasivam⁴, Durga Pokharel⁵, Himabindu Gali⁶, Marina A. Bellani¹, Michael M Seidman¹

¹Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, ²Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, Hunan University, ³Department of Molecular Biology and Genetics, Cornell University, ⁴Department of Cellular and Molecular Medicine, University of Copenhagen, ⁵Horizon Discovery, ⁶Boston University School of Medicine

Summary

While replication fork collisions with DNA adducts can induce double strand breaks, less is known about the interaction between replisomes and blocking lesions. We have employed the proximity ligation assay to visualize these encounters and to characterize the consequences for replisome composition.

Explore More Videos

Replisome Visualization

Antigen tagged DNA

Blocking Lesion

Immunodetection

Cell Cycle Interactions

Ishani Majumdar

Seidman Laboratory

Cell Culture Protocol

Cytoskeleton Extraction Buffer

Formaldehyde Fixation

Triton X 100 Treatment

Chapters in this video

0:04

Introduction

0:51

Cell Preparation

3:58

Proximity Ligation Assay (PLA)

6:20

Results: Representative Replisome Encounter Visualization

7:52

Conclusion

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