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Literature DB >> 28481515

Sequence-Directed Covalent Protein-DNA Linkages in a Single Step Using HUH-Tags.

Klaus N Lovendahl¹, Amanda N Hayward¹, Wendy R Gordon¹.

Abstract

We present a robust strategy to covalently link proteins and DNA using HUH-endonuclease domains as fusion partners (HUH-tags). We show that HUH-tags react robustly with specific sequences of unmodified single-stranded DNA, and we have identified five tags that react orthogonally with distinct DNA sequences. We demonstrate the versatility of HUH-tags as fusion partners in Cas9-mediated gene editing and the construction of doubly DNA-tethered proteins for single-molecule studies. Finally we demonstrate application to cellular imaging in live and fixed cells.

Entities: CellLine Chemical Disease Gene Species

Mesh：

Substances：

Year: 2017 PMID： 28481515 PMCID： PMC5517037 DOI： 10.1021/jacs.7b02572

Source DB: PubMed Journal: J Am Chem Soc ISSN： 0002-7863 Impact factor: 15.419

32 in total

Review 1. Choosing an effective protein bioconjugation strategy.

Authors: Nicholas Stephanopoulos; Matthew B Francis
Journal: Nat Chem Biol Date: 2011-11-15 Impact factor: 15.040

2. Plasmid replication initiator RepB forms a hexamer reminiscent of ring helicases and has mobile nuclease domains.

Authors: D Roeland Boer; José A Ruíz-Masó; José R López-Blanco; Alexander G Blanco; Mireia Vives-Llàcer; Pablo Chacón; Isabel Usón; F Xavier Gomis-Rüth; Manuel Espinosa; Oscar Llorca; Gloria del Solar; Miquel Coll
Journal: EMBO J Date: 2009-06-03 Impact factor: 11.598

Sequence-Directed Covalent Protein-DNA Linkages in a Single Step Using HUH-Tags.

Review 1. Choosing an effective protein bioconjugation strategy.

2. Plasmid replication initiator RepB forms a hexamer reminiscent of ring helicases and has mobile nuclease domains.

3. Spatial control of membrane receptor function using ligand nanocalipers.

4. Multi-enzyme complexes on DNA scaffolds capable of substrate channelling with an artificial swinging arm.

5. Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.

6. Direct observation of individual endogenous protein complexes in situ by proximity ligation.

7. Sequence-specific binding of DNA and RNA to immobilized Nickel ions.

8. Myosin lever arm directs collective motion on cellular actin network.

9. Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT.

10. Barcoding cells using cell-surface programmable DNA-binding domains.

1. Enhanced Molecular Tension Sensor Based on Bioluminescence Resonance Energy Transfer (BRET).

Review 2. Nucleic Acid-Barcoding Technologies: Converting DNA Sequencing into a Broad-Spectrum Molecular Counter.

3. Crystal structure of the Wheat dwarf virus Rep domain.

4. Protein-Nucleic Acid Conjugation with Sterol Linkers Using Hedgehog Autoprocessing.

5. Programmable Assembly of Adeno-Associated Virus-Antibody Composites for Receptor-Mediated Gene Delivery.

6. Stretching DNA to twice the normal length with single-molecule hydrodynamic trapping.

7. Live cell PNA labelling enables erasable fluorescence imaging of membrane proteins.

8. Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template.

Review 9. DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions.

10. Rational design of a DNA sequence-specific modular protein tag by tuning the alkylation kinetics.