Literature DB >> 33739627

Construction and Quantitation of a Selectable Protein Splicing Sensor Using Gibson Assembly and Spot Titers.

Daniel Woods1, Danielle S LeSassier2, Ikechukwu Egbunam3, Christopher W Lennon4.   

Abstract

Inteins (intervening proteins) are translated within host proteins and removed through protein splicing. Conditional protein splicing (CPS), where the rate and accuracy of splicing are highly dependent on environmental cues, has emerged as a novel form of post-translational regulation. While CPS has been demonstrated for several inteins in vitro, a comprehensive understanding of inteins requires tools to quantitatively monitor their activity within the cellular context. Here, we describe a method for construction of a splicing-dependent system that can be used to quantitatively assay for conditions that modulate protein splicing.
© 2021 Wiley Periodicals LLC. Basic Protocol 1: Construction of an intein-containing KanR2 library using Gibson assembly Basic Protocol 2: Phenotype determination using quantitative spot titers Support Protocol 1: Preparation of LB agar plates for spot titers Support Protocol 2: Preparation and transformation of competent M. smegmatis cells. © 2021 Wiley Periodicals LLC.

Entities:  

Keywords:  conditional protein splicing; inteins; mycobacteria

Mesh:

Substances:

Year:  2021        PMID: 33739627      PMCID: PMC7988948          DOI: 10.1002/cpz1.82

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  31 in total

1.  Directed evolution of ligand dependence: small-molecule-activated protein splicing.

Authors:  Allen R Buskirk; Yi-Ching Ong; Zev J Gartner; David R Liu
Journal:  Proc Natl Acad Sci U S A       Date:  2004-07-09       Impact factor: 11.205

2.  Salt-Dependent Conditional Protein Splicing of an Intein from Halobacterium salinarum.

Authors:  Julie N Reitter; Christopher E Cousin; Michael C Nicastri; Mario V Jaramillo; Kenneth V Mills
Journal:  Biochemistry       Date:  2016-02-26       Impact factor: 3.162

3.  Inteins.

Authors:  Christopher W Lennon; Marlene Belfort
Journal:  Curr Biol       Date:  2017-03-20       Impact factor: 10.834

4.  Salt-inducible Protein Splicing in cis and trans by Inteins from Extremely Halophilic Archaea as a Novel Protein-Engineering Tool.

Authors:  Annika Ciragan; A Sesilja Aranko; Igor Tascon; Hideo Iwaï
Journal:  J Mol Biol       Date:  2016-10-06       Impact factor: 5.469

5.  Directed evolution of a small-molecule-triggered intein with improved splicing properties in mammalian cells.

Authors:  Sun H Peck; Irwin Chen; David R Liu
Journal:  Chem Biol       Date:  2011-05-27

6.  Laboratory maintenance of Mycobacterium smegmatis.

Authors:  Anil Kumar Singh; Jean-Marc Reyrat
Journal:  Curr Protoc Microbiol       Date:  2009-08

7.  Mycobacterial recombineering.

Authors:  Julia C van Kessel; Graham F Hatfull
Journal:  Methods Mol Biol       Date:  2008

8.  Small molecule-triggered Cas9 protein with improved genome-editing specificity.

Authors:  Kevin M Davis; Vikram Pattanayak; David B Thompson; John A Zuris; David R Liu
Journal:  Nat Chem Biol       Date:  2015-04-06       Impact factor: 15.040

9.  Spliceosomal Prp8 intein at the crossroads of protein and RNA splicing.

Authors:  Cathleen M Green; Zhong Li; Aaron D Smith; Olga Novikova; Valjean R Bacot-Davis; Fengshan Gao; Saiyang Hu; Nilesh K Banavali; Dennis J Thiele; Hongmin Li; Marlene Belfort
Journal:  PLoS Biol       Date:  2019-10-10       Impact factor: 8.029

Review 10.  Protein splicing: how inteins escape from precursor proteins.

Authors:  Kenneth V Mills; Margaret A Johnson; Francine B Perler
Journal:  J Biol Chem       Date:  2014-04-02       Impact factor: 5.157

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  2 in total

1.  Reactive Chlorine Species Reversibly Inhibit DnaB Protein Splicing in Mycobacteria.

Authors:  Christopher W Lennon; Daniel Wahl; J R Goetz; Joel Weinberger
Journal:  Microbiol Spectr       Date:  2021-09-22

Review 2.  Intein Inhibitors as Novel Antimicrobials: Protein Splicing in Human Pathogens, Screening Methods, and Off-Target Considerations.

Authors:  Diana A Wall; Seanan P Tarrant; Chunyu Wang; Kenneth V Mills; Christopher W Lennon
Journal:  Front Mol Biosci       Date:  2021-10-07
  2 in total

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