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J. Biol. Chem., Vol. 279, Issue 26, 27098-27107, June 25, 2004

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Transcriptional Regulation of Acetylcholinesterase-associated Collagen ColQ

DIFFERENTIAL EXPRESSION IN FAST AND SLOW TWITCH MUSCLE FIBERS IS DRIVEN BY DISTINCT PROMOTERS^*

Henry H. C. Lee, Roy C. Y. Choi¶, Annie K. L. Ting, Nina L. Siow||, Joy X. S. Jiang, Jean Massoulié**, and Karl W. K. Tsim, Held a visiting professorship at Ecole Normale Supérieure in 2001 and 2003

From the Departments of Biology and Molecular Neuroscience Center, The Hong Kong University of Science and Technology, Clear Water Bay Road, Hong Kong SAR, China and ^**Laboratoire de Neurobiologie Cellulaire et Moléculaire, CNRS UMR 8544, Ecole Normale Supérieure, 75005 Paris France

The presence of a collagenous protein (ColQ) characterizes the collagen-tailed forms of acetylcholinesterase and butyrylcholinesterase at vertebrate neuromuscular junctions which is tethered in the synaptic basal lamina. ColQ subunits, differing mostly by their signal sequences, are encoded by transcripts ColQ-1 and ColQ-1a, which are differentially expressed in slow and fast twitch muscles in mammals. Two distinct promoters, pColQ-1 and pColQ-1a, were isolated from the upstream sequences of human COLQ gene; they showed muscle-specific expression and were activated by myogenic transcriptional elements in cultured myotubes. After in vivo DNA transfection, pColQ-1 showed strong activity in slow twitch muscle (e.g. soleus), whereas pColQ-1a was preferably expressed in fast twitch muscle (e.g. tibialis). Mutation analysis of the ColQ promoters suggested that the muscle fiber type-specific expression pattern of ColQ transcripts were regulated by a slow upsteam regulatory element (SURE) and a fast intronic regulatory element (FIRE). These regulatory elements were responsive to a calcium ionophore and to calcineurin inhibition by cyclosporine A. The slow fiber type-specific expression of ColQ-1 was abolished by the mutation of an NFAT element in pColQ-1. Moreover, both the ColQ promoters contained N-box element that was responsible for the synapse-specific expression of ColQ transcripts. These results explain the specific expression patterns of collagen-tailed acetylcholinesterase in slow and fast muscle fibers.

Received for publication, March 8, 2004 , and in revised form, April 16, 2004.

^* This work was supported by the Research Grants Council of Hong Kong (HKUST 6098/02M and 6283/03M) (to K. W. K. T.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this study.

^¶ Supported by a post-doctoral matching fund from Hong Kong University of Science and Technology.

^|| Holds a Croucher Foundation Scholarship.

To whom correspondence should be addressed: Dept. of Biology, The Hong Kong University of Science and Technology, Clear Water Bay Rd., Kowloon, Hong Kong SAR, China. Tel.: 852-2358-7332; Fax: 852-2358-1559; E-mail: botsim{at}ust.hk.

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