Phosphate coordination and movement of DNA in the Tn5 synaptic complex: role of the (R)YREK motif

Journal Article · Thu Jan 15 04:00:00 EST 2009 · Nucleic Acids Res.

DOI:https://doi.org/10.1093/nar/gkn577· OSTI ID:1006916

Klenchin, Vadim A; Czyz, Agata; Goryshin, Igor Y; Gradman, Richard; Lovell, Scott; Rayment, Ivan; Reznikoff, William S ^[1]

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1. UW

Bacterial DNA transposition is an important model system for studying DNA recombination events such as HIV-1 DNA integration and RAG-1-mediated V(D)J recombination. This communication focuses on the role of protein-phosphate contacts in manipulating DNA structure as a requirement for transposition catalysis. In particular, the participation of the nontransferred strand (NTS) 5' phosphate in Tn5 transposition strand transfer is analyzed. The 5' phosphate plays no direct catalytic role, nonetheless its presence stimulates strand transfer {approx}30-fold. X-ray crystallography indicates that transposase-DNA complexes formed with NTS 5' phosphorylated DNA have two properties that contrast with structures formed with complexes lacking the 5' phosphate or complexes generated from in-crystal hairpin cleavage. Transposase residues R210, Y319 and R322 of the (R)YREK motif coordinate the 5' phosphate rather than the subterminal NTS phosphate, and the 5' NTS end is moved away from the 3' transferred strand end. Mutation R210A impairs the 5' phosphate stimulation. It is posited that DNA phosphate coordination by R210, Y319 and R322 results in movement of the 5' NTS DNA away from the 3'-end thus allowing efficient target DNA binding. It is likely that this role for the newly identified RYR triad is utilized by other transposase-related proteins.

• Research Organization: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
• Sponsoring Organization: USDOE
• OSTI ID: 1006916
• Journal Information: Nucleic Acids Res., Journal Name: Nucleic Acids Res. Journal Issue: (18) ; 09, 2008 Vol. 36; ISSN 0305-1048; ISSN NARHAD
• Country of Publication: United States
• Language: ENGLISH

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
CATALYSIS
CLEAVAGE
COMMUNICATIONS
COMPLEXES
COORDINATES
CRYSTALLOGRAPHY
DNA
MUTATIONS
PHOSPHATES
PROTEINS
RECOMBINATION
RESIDUES
STIMULATION
TARGETS