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J. Biol. Chem., Vol. 276, Issue 6, 4373-4381, February 9, 2001

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The Mu Enhancer Is Functionally Asymmetric Both in cis and in trans
TOPOLOGICAL SELECTIVITY OF Mu TRANSPOSITION IS ENHANCER-INDEPENDENT^*

Hong Jiang and Rasika M. Harshey

From the Section of Molecular Genetics and Microbiology and Institute of Cellular and Molecular Biology, University of Texas, Austin, Texas 78712

Mu DNA transposition from a negatively supercoiled DNA substrate requires interaction of an enhancer element with the left (attL) and right (attR) ends of Mu. The orientation of the L and R ends with respect to each other (inverted) and with respect to the enhancer is normally inviolate. We show that when the enhancer is provided in trans as a linear fragment, the head to head orientation of the L/R ends is still required. Each functional half of the linear enhancer maintains the same "cross-wise" interaction with the subsites L1 and R1, when present in cis or in trans. In reactions catalyzed by an enhancer-independent variant of the Mu transposase, the need for negative supercoiling of the substrate and the inverted orientation of L and R ends is not relaxed. These results show that the orientation specificity of the enhancer is not determined by its topological linkage to the Mu ends. There is a functional asymmetry inherent to the enhancer. Furthermore, the enhancer does not directly impose topological constraints on the transposition reaction or specify the reactive orientation of the Mu ends.

This article has been cited by other articles:

				Z. Yin and R. M. Harshey Enhancer-independent Mu transposition from two topologically distinct synapses PNAS, December 27, 2005; 102(52): 18884 - 18889. [Abstract] [Full Text] [PDF]