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A more recent version of this article appeared on June 1, 2005
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Submitted on October 28, 2004
Revised on March 3, 2005
Accepted on March 21, 2005
*Comparative Cell Biology Group, Temasek Life Science Laboratory, National University of Singapore, Singapore 117604, Singapore; Laboratory of Plant Molecular Biology and Bio-Imaging Resource Center, The Rockefeller University, New York, NY 10021
Monitoring Editor: David Drubin
The Woronin body (WB) is a peroxisome-related organelle that is centered on a crystalline core of the HEX-1 protein, which functions to seal septal pores of filamentous ascomycetes in response to cellular damage. Here we investigate the cellular and genetic control of WB-formation and show that polarized hex-1 gene expression determines WB-biogenesis at the growing hyphal apex. We find that intron splicing is coupled to efficient hex-1 gene expression and strikingly, when the yellow fluorescent protein (YFP) was expressed from hex-1 regulatory sequences; we observed a fluorescent gradient that was maximal in apical cells. Moreover, endogenous hex-1 transcripts were specifically enriched at the leading edge of the fungal colony, while other transcripts accumulated in basal regions. Time-lapse confocal microscopy showed that HEX-1 crystals normally formed in the vicinity of the hyphal apex in large peroxisomes, which matured and were immobilized at the cell periphery as cells underwent septation. When the hex-1 structural gene was expressed from regulatory sequences of an abundant, basally localized transcript, WB-core formation was redetermined to basal regions of the colony and these strains displayed loss-of-function phenotypes specifically in apical hyphal compartments. These results show that localized gene expression is a key determinant of spatially restricted WB-assembly. We speculate that this type of regulation constitutes a widely utilized mechanism for determining cellular activity in specific regions of the fungal hypha.
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