tropic growth is facilitated by polarized protein complexes tan truong biol 503 may 4, 2010
Post on 23-Jan-2016
218 views
TRANSCRIPT
Tropic growth is facilitated by polarized protein complexes
Tan TruongBIOL 503
May 4, 2010
• Galvanotropism – growth in response to external electric field • Thigmotropism – growth in response to contact• Chemotropism – growth in response to chemicals• Geotropism/gravitropism – growth in response to gravity• Heliotropism/Phototropism – growth in response to sunlight• Hydrotropism – growth in response to water• Thermotropism – growth in response to temperature changes• Specific Host Tropism (amphotropism, neurotropism, etc.)
Tropism
Galvanotropism
C. albicans hyphal reorientation upon applied current
(Brand et al., 2007)
Thigmotropism
C. albicans hyphal reorientation upon surface ridge contact
(Brand et al., 2007)
Neurospora crassa chemotropic attraction (Mak-2 localizing to CAT)
Chemotropism
(Gleissner et al., 2009)
Phycomyces exhibiting phototropism
Phototropism
(Bergman et al., 1969)
• Candida albicans - thin-walled, small budding yeast
• Saccharomyces cerevisiae – model organism
• Candida glabrata – non-dimorphic, haploid yeast; opportunistic pathogen; Candidemia
Fungal Species
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
(Brand & Gow, 2009)
Outline
• Environmental cues can induce tropism
• Environmental cues can promote the uptake of other environmental cues, which can induce directional growth
• Strategically-positioned receptors can facilitate tropism
Hypotheses
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Ozeki-Miyawaki et al., 2005)
Stretch described by (Murase et al., 2001)
Figure 1: S. cerivisiae Mid1 is a stretch-activated Ca2+ channel
Figure 2: Extracellular Ca2+ affects cathodal emergence of C. albicans hyphae
(Brand et al., 2007)
Figure 3: Thigmotropic response is attenuated in C. albicans Ca2+–signaling –pathway mutants
(Brand et al., 2007)
(Brand et al., 2007)
Figure 4: Proposed model for Ca2+ uptake in galvanotropic and thigmotropic growth
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Miyazaki et al., 2010)
Figure 5: Calcineurin is important for C. glabrata azole resistance
Cnb1 = Calcineurin B subunit
(Karababa et al., 2006)
Figure 6: Calcineurin & Crz1 regulate the expression of C. albicans Ca2+-dependent genes
Cna = Calcineurin A subunit
(Brand & Gow, 2009)
Re-cap
(Kang et al., 2001)
Figure 7: S. cerevisiae Bud5 interacts with Axl2
S10 = soluble fraction
a = haploid yeasta/α = diploid yeastb = untagged protein
b b
(Marston et al., 2001)
Figure 8: Bud5 localizes at pre-bud sites in both haploid and diploid S. cerevisiae cells
Bright field im
agesha
ploi
ddi
ploi
d
(Marston et al., 2001)
Figure 9: S. cerevisiae Bud2 & Bud5 localize at pre-bud sites
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
Figure 10: Rsr1 & Bud2 are important for actin patch polarization in C. albicans
(Hausauer et al., 2005)
Stained with Alexa phalloidin (actin)DIC microscopy
Figure 11: C. albicans Rsr1 & Bud2 are important for hyphal morphogenesis
(Hausauer et al., 2005)
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
Figure 12: Yeast Two-Hybrid (Y2H) Screening
http://en.wikipedia.org/wiki/Two-hybrid_screening
(Barale et al., 2004)
Figure 13: S. cerevisiae Cdc24 interacts with both Far1 & Bem1
(Brand & Gow, 2009)
(Barale et al., 2004)
Figure 14: S. cerevisiae Cdc24 responds to α-factor
(Brand & Gow, 2009)
Re-cap
GST-
(Mionnet et al., 2008)
Figure 15: S. cerevisiae Cdc24 associates with Cdc42
(Brand & Gow, 2009)
Re-cap
(Brand & Gow, 2009)
Re-cap
(B)
(A)
(Oehlen et al., 1996)
Figure 16: S. cerevisiae Ste2 regulates far1
(Brand & Gow, 2009)
Conclusion
• Environmental cues can induce tropism
• Environmental cues can promote the uptake of other environmental cues, which can induce directional growth
• Strategically-positioned receptors can facilitate tropism
Take-Home Messages
?
(Brand & Gow, 2009)
(Brand et al., 2007)
Proposed model for Ca2+ uptake in galvanotropic and thigmotropic growth
Tropic growth is facilitated by polarized protein complexes
Tan TruongBIOL 503
May 4, 2010
?¿
BibliographyBarale, S., McCusker, D., Arkowitz, R.A. (2004) The exchange factor Cdc24 is required for cell fusion during yeast mating. Eukaryotic Cell 3, 1049-61
Bergman, K., Burke, P., Cerda, E., David, C., Delbruck, M., Foster, W., Goodel, E., Heisenberg, M., Meissner, G., Zakokar, M., Shropshire, W. (1969) Phycomyces. American Society for Microbiology 33, 99-157
Brand, A., Gow, N. (2009) Mechanisms of hypha orientation of fungi. Science Direct 12, 350-357
Brand, A., Shanks, S., Duncan, V., Yang, M., Mackenzie, K., Gow, N. (2007) Hyphal Orientation of Candida albicans is Regulated by a Calcium Dependent Mechanism. Current Biology 16, 347-352
Brand, A., Vacharaksa, A., Bendel, C., Norton, J., Haynes, P., Henry-Stanley, M., Wells, C., Ross, K., Gow, N.A.R., Gale, C.A. (2008) An Internal Polarity Landmark Is Important for Externally Induced Hyphal Behaviors in Candida albicans. Eukaryotic Cell 7, 712-20
Casamayor, A., Snyder, M. (2002) Bud-site selection and cell polarity in budding yeast. Current Opinion in Microbiology 5, 179-86
Davies, J.M., Stacey, A.J., Gilligan, C.A. (1999) Candida albicans hyphal invasion: thigmotropism or chemotropism? FEMS Microbiology Letters 171, 245-9
Flesslssner, A., Leeder, A., Roca, M., Read, N., Glass, N. (2009) Oscillatory recruitment of signaling proteins to cell tips promotes coordinated behavior during cell fusion. PNAS 106, 19387-19392
Hasan, S., Delfosse, E.S., Aracil, E., Lewis, R.C. (1992) Host-specificity of Uromyces heliotropii, a fungal agent for the biological control of common heliotrope (Heliotropium europaeum) in Australia. Annals of Applied Biology 121, 697 - 705
Hausauer, D.L., Gerami-Nejad, M., Kristler-Anderson, C., Gale, C.A. (2005) Hyphal Guidance and Invasive Growth in Candida albicans Require the Ras-Like GTPase Rsr1p and Its GTPase-Activating Protein Bud2p. Eukaryotic Cell 7, 1273-1286
Hida, H., Nakamura, H., Ono, T., Okumura, M., Anraku, Y. (1994) MID1, a Novel Saccharomyces cerevisiae Gene Encoding a Plasma Membrane Protein Is Required for Ca2+ Influx and Mating. Molecular and Cell biology 14, 8259-8271
Kang, P.J., Sanson, A., Bongyong, L., Park, H.O. (2001) A GDP/GTP Exchange Factor Involved in Linking a Spatial Landmark to Cell Polarity. Science 292, 1376 – 1378
Karababa, M., Valentino, E., Pardini, G., Coste, A.T., Bille, J., Sanglard, D. (2006) CRZ1, a target of the calcineurin pathway in Candida albicans. Molecular Microbiology 59, 1429-51
Marston, A.L., Chen, T., Yang, M.C., Belhumeur, P., Chant, J. (2001) A localized GTPase exchange factor, Bud5, determines the orientation of division axes in yeast. Current Biology 11, 803–807
Mionnet, C., Bogliolo, S., Arkowitz, R. (2008) Oligomerization Regulates the Localization of Cdc24, the Cdc42 Activator in Saccharomyces cerevisiae. The Journal of Biological Chemistry 283, 17515-17530
Murase, K., Naruse, K., Kimura, A., Okumura, K., Hayakawa, T., Sokabe, M. (2001) Protamine augments stretch induced calcium increase in vascular endothelium. Journal of Pharmacology 134, 1403-1410
Oehlen L.J.W.M., Mckinney, J.D., Cross, F.R. (1996) The Exchange Factor Cdc24 Is Required for Cell Fusion during Yeast Mating. Eukaryotic Cell 11, 1049–1061
Ozeki-Miyawaki, C., Moriya, Y., Tatsumi, H., Iida, H., Sokabe, M. (2005) Identification of functional domains of Mid1, a stretch-activated channel component, necessary for localization to the plasma membrane and Ca2+ permeation. Experimental Cell Research 311, 84-85.
Sbrana, C., Giovannetti, M. (2005) Chemotropism in the arbuscular mycorrhizal fungus Glomus mosseae. Mycorrhizae 15, 539-45
Schimek, C., Eibel, P., Grolig, F., Horie, T., Ootaki, T., Galland, P. (1999) Gravitropism in Phycomyces: a role for sedimenting protein crystals and floating lipid globules. Planta 210, 132-42
Watts, H.J., Véry, A.A., Perera, T.H., Davies, J.M., Gow, N.A. (1998) Thigmotropism and stretch-activated channels in the pathogenic fungus Candida albicans. Microbiology 144, 689-95
Zhou, X.L., Stumpf, M.A., Hoch, H.C., Kung, C. (1991) Mechanosensitive Channel in Whole Cells and in Membrane Patches of the Fungus Uromyces. Science 15, 85-92
Bibliography
Any Questions?