scholarly journals SIRF: Quantitative in situ analysis of protein interactions at DNA replication forks

2018 ◽  
Vol 217 (4) ◽  
pp. 1553-1553 ◽  
Author(s):  
Sunetra Roy ◽  
Jessica W. Luzwick ◽  
Katharina Schlacher
Keyword(s):  
Dna Replication ◽  
Protein Interactions ◽  
In Situ Analysis ◽  
Replication Forks

scholarly journals SIRF: Quantitative in situ analysis of protein interactions at DNA replication forks

2018 ◽  
Vol 217 (4) ◽  
pp. 1521-1536 ◽  
Author(s):  
Sunetra Roy ◽  
Jessica W. Luzwick ◽  
Katharina Schlacher
Keyword(s):  
Quantitative Analysis ◽  
Dna Replication ◽  
Protein Interactions ◽  
Accelerated Aging ◽  
In Situ Analysis ◽  
Replication Forks ◽  
Cancer Etiology ◽  
Validation Data ◽  
Stalled Replication Forks

DNA replication reactions are central to diverse cellular processes including development, cancer etiology, drug treatment, and resistance. Many proteins and pathways exist to ensure DNA replication fidelity and protection of stalled or damaged replication forks. Consistently, mutations in proteins involved in DNA replication are implicated in diverse diseases that include defects during embryonic development and immunity, accelerated aging, increased inflammation, blood disease, and cancer. Thus, tools for efficient quantitative analysis of protein interactions at active and stalled replication forks are key for advanced and accurate biological understanding. Here we describe a sensitive single-cell–level assay system for the quantitative analysis of protein interactions with nascent DNA. Specifically, we achieve robust in situ analysis of protein interactions at DNA replication forks (SIRF) using proximity ligation coupled with 5′-ethylene-2′-deoxyuridine click chemistry suitable for multiparameter analysis in heterogeneous cell populations. We provide validation data for sensitivity, accuracy, proximity, and quantitation. Using SIRF, we obtained new insight on the regulation of pathway choice by 53BP1 at transiently stalled replication forks.

scholarly journals SIRFing the replication fork: Assessing protein interactions with nascent DNA

2018 ◽  
Vol 217 (4) ◽  
pp. 1177-1179
Author(s):  
Dana Branzei ◽  
Michele Giannattasio
Keyword(s):  
Quantitative Analysis ◽  
Dna Replication ◽  
Single Cell ◽  
Protein Interactions ◽  
Replication Fork ◽  
Cell Populations ◽  
Replication Forks ◽  
Multiparameter Measurements ◽  
Stalled Replication Forks

Roy et al. (2018. J. Cell. Biol. https://doi.org/10.1083/jcb.201709121) describe an ingenious single-cell assay system, in situ analysis of protein interactions at DNA replication forks (SIRF), for the quantitative analysis of protein interactions with nascent DNA at active and stalled replication forks. The sensitive and accurate SIRF methodology is suitable for multiparameter measurements in cell populations.

KISS: A Mammalian Two-Hybrid Method for In Situ Analysis of Protein–Protein Interactions

2018 ◽  
pp. 269-278 ◽  
Author(s):  
Delphine Masschaele ◽  
Sarah Gerlo ◽  
Irma Lemmens ◽  
Sam Lievens ◽  
Jan Tavernier
Keyword(s):  
Protein Interactions ◽  
Hybrid Method ◽  
In Situ Analysis ◽  
Protein Protein Interactions ◽  
Two Hybrid

Fine Structural in Situ Analysis of Nascent DNA Movement Following DNA Replication

2000 ◽  
Vol 260 (2) ◽  
pp. 313-323 ◽  
Author(s):  
Françoise Jaunin ◽  
Astrid E. Visser ◽  
Dusan Cmarko ◽  
Jacob A. Aten ◽  
Stanislav Fakan
Keyword(s):  
Dna Replication ◽  
In Situ Analysis

In Situ Analysis and Visualization with CREATE-AVTMHelios

2021 ◽  
Author(s):  
Andrew C. Bauer ◽  
James R. Forsythe ◽  
Jay Sitaraman ◽  
Andrew M. Wissink ◽  
Buvaneswari Jayaraman ◽  
...  
Keyword(s):  
In Situ Analysis
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