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Science 3 August 1990:
Vol. 249. no. 4968, pp. 511 - 517
DOI: 10.1126/science.2166334
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Articles

Science, Vol 249, Issue 4968, 511-517
Copyright © 1990 by American Association for the Advancement of Science

articles

The Hin invertasome: protein-mediated joining of distant recombination sites at the enhancer

KA Heichman and RC Johnson

Department of Biological Chemistry, UCLA School of Medicine.

The Hin protein binds to two cis-acting recombination sites and catalyzes a site-specific DNA inversion reaction that regulates the expression of flagellin genes in Salmonella. In addition to the Hin protein and the two recombination sites that flank the invertible segment, a third cis-acting recombinational enhancer sequence and the Fis protein, which binds to two sites within the enhancer, are required for efficient recombination. Intermediates of this reaction were trapped during DNA strand cleavage and analyzed by gel electrophoresis and electron microscopy in order to determine their structure and composition. The analyses demonstrate that the recombination sites are assembled at the enhancer into a complex nucleo-protein structure (termed the invertasome) with the looping of the three segments of intervening DNA. Antibody studies indicated that Fis physically interacts with Hin and that both proteins are intimately associated with the invertasome. In order to achieve this protein-protein interaction and assemble the invertasome, the substrate DNA must be supercoiled.


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F. G. Albert, E. C. Bronson, D. J. Fitzgerald, and J. N. Anderson (1995)
J. Biol. Chem. 270, 23570-23581
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Hin recombinase bound to DNA: the origin of specificity in major and minor groove interactions.
J. Feng, R. Johnson, and R. Dickerson (1994)
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Analysis of a nucleoprotein complex: the synaptosome of gamma delta resolvase.
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Configuration of DNA strands and mechanism of strand exchange in the Hin invertasome as revealed by analysis of recombinant knots..
K A Heichman, I P Moskowitz, and R C Johnson (1991)
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Alignment of recombination sites in Hin-mediated site-specific DNA recombination..
I P Moskowitz, K A Heichman, and R C Johnson (1991)
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Sequence-specific DNA binding by the c-Myc protein.
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J. Biol. Chem. 276, 4373-4381
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