Hin recombinase
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Hin recombinase is a 21kD protein composed of 198 amino acids that is found in the bacteria Salmonella. Hin belongs to the serine recombinase family of DNA invertases in which it relies on the active site serine to initiate DNA cleavage and recombination. The related protein, gamma-delta resolvase shares high similarity to Hin, of which much structural work has been done, including structures bound to DNA and reaction intermediates. Hin functions to invert a 900bp DNA segment within the salmonella genome that contains a promoter for downstream flagellar genes, fljA and fljB. Inversion of the intervening DNA alternates the direction of the promoter and thereby alternates expression of the flagellar genes. This is advantageous to the bacterium as a means of escape from the host immune response.
Hin functions by binding to two 26bp imperfect inverted repeat sequences as a homodimer. These hin binding sites flank the invertible segment which not only encodes the Hin gene itself, but also contains an enhancer element to which the bacterial Fis proteins binds with nanomolar affinity. Four molecules of Fis bind to this site as a homodimers and are required for the recombination reaction to proceeded.
The initial reaction requires binding of Hin and Fis to their respective DNA sequences and assemble into a higher-order nucleoprotein complex with branched plectonemic supercoils with the aid of the DNA bending protein HU. At this point, it is believed that the Fis protein modulates subtle contacts to activate the reaction, possibly through direct interactions with the Hin protein. Activation of the 4 catalytic serine residues within the Hin tetramer make a 2-bp double stranded DNA break and forms a covalent reaction intermediate. The DNA cleavage event also requires the divalent metal cation magnesium. A large conformational change reveals a large hydrophobic interface that allows for subunit rotation which may be driven by superhelical torsion within the protein-DNA complex. Subsequent to a single 180° rotation, Hin returns to its native conformation and relegates the cleaved DNA, without the aid of high energy cofactors and without the loss of any DNA.
[edit] References
- Dhar G, Sanders E, Johnson R (2004). "Architecture of the hin synaptic complex during recombination: the recombinase subunits translocate with the DNA strands". Cell 119 (1): 33–45. doi:. PMID 15454079.
- Sanders E, Johnson R (2004). "Stepwise dissection of the Hin-catalyzed recombination reaction from synapsis to resolution". J Mol Biol 340 (4): 753–66. doi:. PMID 15223318.
- Kamtekar S, Ho R, Cocco M, Li W, Wenwieser S, Boocock M, Grindley N, Steitz T (2006). "Implications of structures of synaptic tetramers of gamma delta resolvase for the mechanism of recombination". Proc Natl Acad Sci U S A 103 (28): 10642–7. doi:. PMID 16807292.
- Li W, Kamtekar S, Xiong Y, Sarkis G, Grindley N, Steitz T (2005). "Structure of a synaptic gammadelta resolvase tetramer covalently linked to two cleaved DNAs". Science 309 (5738): 1210–5. doi:. PMID 15994378.
