Eukaryotic Cell
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Eukaryotic Cell, October 2007, p. 1824-1840, Vol. 6, No. 10
1535-9778/07/$08.00+0     doi:10.1128/EC.00188-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Role of Actin Cytoskeletal Dynamics in Activation of the Cyclic AMP Pathway and HWP1 Gene Expression in Candida albicans{triangledown} ,{dagger}

Michael J. Wolyniak and Paula Sundstrom*

Microbiology and Molecular Pathogenesis Program, Dartmouth Medical School, Hanover, New Hampshire 03755

Received 23 May 2007/ Accepted 7 August 2007

Changes in gene expression during reversible bud-hypha transitions of the opportunistic fungal pathogen Candida albicans permit adaptation to environmental conditions that are critical for proliferation in host tissues. Our previous work has shown that the hypha-specific adhesin gene HWP1 is up-regulated by the cyclic AMP (cAMP) signaling pathway. However, little is known about the potential influences of determinants of cell morphology on HWP1 gene expression. We found that blocking hypha formation with cytochalasin A, which destabilizes actin filaments, and with latrunculin A, which sequesters actin monomers, led to a loss of HWP1 gene expression. In contrast, high levels of HWP1 gene expression were observed when the F-actin stabilizer jasplakinolide was used to block hypha formation, suggesting that HWP1 expression could be regulated by actin structures. Mutants defective in formin-mediated nucleation of F-actin were reduced in HWP1 gene expression, providing genetic support for the importance of actin structures. Kinetic experiments with wild-type and actin-deficient cells revealed two distinct phases of HWP1 gene expression, with a slow, actin-independent phase preceding a fast, actin-dependent phase. Low levels of HWP1 gene expression that appeared to be independent of stabilized actin and cAMP signaling were detected using indirect immunofluorescence. A connection between actin structures and the cAMP signaling pathway was shown using hyper- and hypomorphic cAMP mutants, providing a possible mechanism for up-regulation of HWP1 gene expression by stabilized actin. The results reveal a new role for F-actin as a regulatory agent of hypha-specific gene expression at the bud-hypha transition.

* Corresponding author. Mailing address: Dartmouth Medical School, Vail Building, HB7550, Hanover, NH 03755. Phone: (603) 650-1629. Fax: (603) 650-1318. E-mail: Paula.R.Sundstrom{at}Dartmouth.edu

{triangledown} Published ahead of print on 22 August 2007.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, October 2007, p. 1824-1840, Vol. 6, No. 10
1535-9778/07/$08.00+0     doi:10.1128/EC.00188-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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