A Neurospora crassa morphological mutant showing reduced adenylate cyclase activity

Terenzi, Héctor Francisco; Flawiá, Mirtha M.; Torres, Héctor N.

doi:10.1016/s0006-291x(74)80241-x

Cited by 85 publications

(71 citation statements)

References 14 publications

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“…Similar phenotypes were described also for other adenylyl cyclase mutant strains from filamentous fungi including Magnaporthe grisea, Sclerotinia sclerotiorum, Aspergillus fumigatus, Trichoderma virens and N. crassa (Terenzi et al 1974;Choi and Dean 1997;Liebmann et al 2003;Jurick and Rollins 2007;Mukherjee et al 2007). The addition of dibutyryl-cAMP to the Dsac1 strain of S. macrospora partially restores the growth defect as was previously shown for the close relative N. crassa (Terenzi et al 1974;Rosenberg and Pall 1979). Besides the vegetative phenotype, the deletion of the adenylyl cyclase gene leads to an impairment in fruiting body development.…”

Section: Discussionsupporting

confidence: 76%

Three α-Subunits of Heterotrimeric G Proteins and an Adenylyl Cyclase Have Distinct Roles in Fruiting Body Development in the Homothallic Fungus Sordaria macrospora

et al. 2008

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Sordaria macrospora, a self-fertile filamentous ascomycete, carries genes encoding three different a-subunits of heterotrimeric G proteins (gsa, G protein Sordaria alpha subunit). We generated knockout strains for all three gsa genes (Dgsa1, Dgsa2, and Dgsa3) as well as all combinations of double mutants. Phenotypic analysis of single and double mutants showed that the genes for Ga-subunits have distinct roles in the sexual life cycle. While single mutants show some reduction of fertility, double mutants Dgsa1Dgsa2 and Dgsa1Dgsa3 are completely sterile. To test whether the pheromone receptors PRE1 and PRE2 mediate signaling via distinct Ga-subunits, two recently generated Dpre strains were crossed with all Dgsa strains. Analyses of the corresponding double mutants revealed that compared to GSA2, GSA1 is a more predominant regulator of a signal transduction cascade downstream of the pheromone receptors and that GSA3 is involved in another signaling pathway that also contributes to fruiting body development and fertility. We further isolated the gene encoding adenylyl cyclase (AC) (sac1) for construction of a knockout strain. Analyses of the three DgsaDsac1 double mutants and one Dgsa2Dgsa3Dsac1 triple mutant indicate that SAC1 acts downstream of GSA3, parallel to a GSA1-GSA2-mediated signaling pathway. In addition, the function of STE12 and PRO41, two presumptive signaling components, was investigated in diverse double mutants lacking those developmental genes in combination with the gsa genes. This analysis was further completed by expression studies of the ste12 and pro41 transcripts in wild-type and mutant strains. From the sum of all our data, we propose a model for how different Ga-subunits interact with pheromone receptors, adenylyl cyclase, and STE12 and thus cooperatively regulate sexual development in S. macrospora.

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Section: Discussionsupporting

confidence: 76%

Three α-Subunits of Heterotrimeric G Proteins and an Adenylyl Cyclase Have Distinct Roles in Fruiting Body Development in the Homothallic Fungus Sordaria macrospora

et al. 2008

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“…Since there were only small differences 6dA was used in the subsequent experiments. The cr-1 mutant is adenylate cyclase-deficient and therefore exhibits reduced concentrations of endogenous cAMP (Terenzi et al, 1974). The acon-2 mutant is defective in an early developmental step of conidia formation, the minor constriction budding (for a review see Russo & Pandit, 1992).…”

Section: Methodsmentioning

confidence: 99%

Differential HSC70 expression during asexual development of Neurospora crassa

et al. 1997

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The constitutive and the heat-shock-induced expression of members of heatshock protein families changed during vegetative development and conidiation of Neurospora crassa as determined by two-dimensional gel electrophoresis. Western blot and ELSA analyses revealed the highest amounts of the constitutive heat-shock protein 70 (HSC70) in conidiating aerial hyphae and dormant conidia. During conidial germination the amount of HSC7O decreased and subsequently increased during vegetative growth. Stationary mycelia and young aerial hyphae exhibited the lowest HSC7O level. The stationary-phase-dependent decrease in HSC70 was accompanied by a concomitant increase in i t s nuclear localization, whereas no significant changes in the amount of nuclear HSC7O were found during aerial hyphae development. The CAMP content during aerial hyphae development was inversely correlated with that of HSC7O. To examine possible causal relations between HSC70 expression and CAMP content, the adenylate-cyclase-def icient mutant crisp (cr-I) was analysed, which exhibits low concentrations of endogenous CAMP. This mutant, however, showed a lower constitutive HSC7O level, compared to the bdA strain. Treatment of the bd strain and cr-I mutant with 20 pM 8-bromo-CAMP did not result in significant changes of the constitutive HSC7O level, but in the level of heat-induced HSOHSP70. In a developmental mutant (acon-2) which is defective in a differentiation step toward conidiation, the expression of HSC7O in aerial hyphae was delayed until the first proconidial chains were observed. It is concluded that the differential expression of HSUHSP7O does not depend on different nuclear levels of HSC7O or on changes in CAMP concentrations, but rather on developmental genes controlling conidiation.

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“…Colonial growth and hyper-conidiation of the cr-1 mutant were partially corrected by exogenous cAMP at a concentration of 5 mM, as described previously (Terenzi et al 1974;Rosenberg and Pall 1979). However, the addition of cAMP did not affect the phenotypes of pkac-1 mutants (Fig.…”

Section: Isolation Of Rip-induced Mutants Of Camp-dependentmentioning

confidence: 99%

“…In Neurospora crassa, an adenylate cyclase mutant, cr-1 ( crisp-1 ), lacks a detectable amount of cAMP (Terenzi et al 1974). When the cr-1 mutant is cultured on a solid medium, it grows colonially, lacks appreciable aerial hyphae, and prematurely produces macroconidia directly on the surface of the mycelium.…”

Section: Introductionmentioning

confidence: 99%

A catalytic subunit of cyclic AMP-dependent protein kinase, PKAC-1, regulates asexual differentiation in Neurospora crassa

et al. 2005

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A cyclic AMP (cAMP)-dependent protein kinase pathway has been shown to regulate growth, morphogenesis and virulence in filamentous fungi. However, the precise mechanisms of regulation through the pathway remain poorly understood. In Neurospora crassa , the cr-1 adenylate cyclase mutant exhibits colonial growth with short aerial hyphae bearing conidia, and the mcb mutant, a mutant of the regulatory subunit of cAMP-dependent protein kinase (PKA), shows the loss of growth polarity at the restrictive temperature. In the present study, we isolated mutants of the catalytic subunit of the PKA gene pkac-1 through the process of repeat-induced point mutation (RIP). PKA activity of the mutants obtained through RIP was undetectable. The genome sequence predicts two distinct catalytic subunit genes of PKA, named pkac-1 (NCU06240.1, AAF75276) and pkac-2 (NCU00682.1), as is the case in most filamentous fungi. The results suggest that PKAC-1 works as the major PKA in N. crassa. The phenotype of the pkac-1 mutants included colonial growth, short aerial hyphae, premature conidiation on solid medium, inappropriate conidiation in submerged culture, and increased thermotolerance. This phenotype of pkac-1 mutants resembled to that of cr-1 mutants, except that the addition of cAMP did not rescue the abnormal morphology of pkac-1 mutants. The loss of growth polarity at the restrictive temperature in the mcb mutant was suppressed by pkac-1 mutation. These results suggest that the signal transduction pathway mediated by PKAC-1 plays an important role in regulation of aerial hyphae formation, conidiation, and hyphal growth with polarity.

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A Neurospora crassa morphological mutant showing reduced adenylate cyclase activity

Cited by 85 publications

References 14 publications

Three α-Subunits of Heterotrimeric G Proteins and an Adenylyl Cyclase Have Distinct Roles in Fruiting Body Development in the Homothallic Fungus Sordaria macrospora

Three α-Subunits of Heterotrimeric G Proteins and an Adenylyl Cyclase Have Distinct Roles in Fruiting Body Development in the Homothallic Fungus Sordaria macrospora

Differential HSC70 expression during asexual development of Neurospora crassa

A catalytic subunit of cyclic AMP-dependent protein kinase, PKAC-1, regulates asexual differentiation in Neurospora crassa

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