Coordinated Regulation of Protoperithecium Development by MAP Kinases MAK-1 and MAK-2 in Neurospora crassa

Mitogen-activated protein (MAP) kinase pathways function as signaling hubs that are integral for many essential cellular processes, including sexual development. The molecular mechanisms and cross-talk between PR and CWI MAP kinase pathways have been extensively studied during asexual development. However, if these can be extended to sexual development remains elusive. By analyzing genome-wide transcriptional responses to deletion of each of two MAP kinase coding genes mak-2 (PR-MAP kinase pathway) and mak-1 (CWI-MAP kinase pathway) in Neurospora crassa during protoperithecium formation, 430 genes co-regulated by the MAK-1 and MAK-2 proteins were found, functionally enriched at integral components of membrane and oxidoreductase. These genes include 13 functionally known genes participating in sexual development (app, poi-2, stk-17, fsd-1, vsd-8, and NCU03863) and melanin synthesis (per-1, pkh-1, pkh-2, mld-1, scy-1, trn-2, and trn-1), as well as a set of functionally unknown genes. Phenotypic analysis of deletion mutants for the functionally unknown genes revealed that 12 genes were essential for female fertility. Among them, single-gene deletion mutants for NCU07743 (named as pfd-1), NCU02250 (oli), and NCU05948 (named as pfd-2) displayed similar protoperithecium development defects as the Δmak-1 and Δmak-2 mutants, failing to form protoperithecium. Western blotting analysis showed that both phosphorylated and total MAK-1 proteins were virtually abolished in the Δnrc-1, Δmek-2, and Δmak-2 mutants, suggesting that the posttranscriptional regulation of MAK-1 is dependent on the PR-MAP kinase pathway during the protoperithecium development. Taken together, this study revealed the regulatory roles and cross-talk between PR and CWI-MAP kinase pathways during protoperithecium development.

Download Full-text

Defining a mechanism of circadian output in Neurospora crassa : temporal regulation of a MAP kinase pathway

The FASEB Journal ◽

10.1096/fasebj.25.1_supplement.944.1 ◽

2011 ◽

Vol 25 (S1) ◽

Author(s):

Charles S Goldsmith ◽

Teresa Lamb ◽

Lindsay Bennett ◽

Katelyn Finch ◽

Deborah Bell‐Pedersen

Keyword(s):

Map Kinase ◽

Neurospora Crassa ◽

Temporal Regulation ◽

Map Kinase Pathway ◽

Kinase Pathway

Download Full-text

PtdIns(3)P accumulation in triple lipid-phosphatase-deletion mutants triggers lethal hyperactivation of the Rho1p/Pkc1p cell-integrity MAP kinase pathway

Journal of Cell Science ◽

10.1242/jcs.02649 ◽

2005 ◽

Vol 118 (23) ◽

pp. 5589-5601 ◽

Cited By ~ 16

Author(s):

W. R. Parrish

Keyword(s):

Map Kinase ◽

Deletion Mutants ◽

Cell Integrity ◽

Lipid Phosphatase ◽

P Accumulation ◽

Map Kinase Pathway ◽

Kinase Pathway

Download Full-text

The Colletotrichum lagenarium Ste12-Like Gene CST1 Is Essential for Appressorium Penetration

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2003.16.4.315 ◽

2003 ◽

Vol 16 (4) ◽

pp. 315-325 ◽

Cited By ~ 75

Author(s):

Gento Tsuji ◽

Satoshi Fujii ◽

Seiji Tsuge ◽

Tomonori Shiraishi ◽

Yasuyuki Kubo

Keyword(s):

Map Kinase ◽

Lipid Droplets ◽

Map Kinases ◽

Cellulose Membrane ◽

Appressorium Formation ◽

Colletotrichum Lagenarium ◽

Protein Map ◽

Mitogen Activated Protein ◽

Map Kinase Pathway ◽

Kinase Pathway

Colletotrichum lagenarium is the causal agent of anthracnose of cucumber. This fungus produces a darkly melanized infection structure, appressoria, to penetrate the host leaves. The C. lagenarium CMK1 gene, a homologue of the Saccharomyces cerevisiae FUS3/KSS1 mitogen-activated protein (MAP) kinase genes, was shown to regulate conidial germination, appressorium formation, and invasive growth. In S. cerevisiae, Ste12p is known to be a transcriptional factor downstream of Fus3p/Kss1p MAP kinases. To evaluate the CMK1 MAP kinase pathway, we isolated the Ste12 homologue CST1 gene from C. lagenarium and characterized. The cst1Δ strains were nonpathogenic on intact host leaves, but could form lesions when inoculated on wounded leaves. Conidia of the cst1Δ strains could germinate and form melanized appressoria on both host leaf surface and artificial cellulose membrane, but could not produce infectious hyphae from appressoria, suggesting that CST1 is essential for appressorium penetration in C. lagenarium. In addition, matured appressoria of the cst1Δ strains contained an extremely low level of lipid droplets compared with that of the wild-type strain. Lipid droplets were abundant in conidia of the cst1Δ strains, but rapidly disappeared during appressorium formation. This misscheduled lipid degradation might be related to the failure of appressorium penetration in the cst1Δ strain.

Download Full-text

WSC-1 and HAM-7 Are MAK-1 MAP Kinase Pathway Sensors Required for Cell Wall Integrity and Hyphal Fusion in Neurospora crassa

PLoS ONE ◽

10.1371/journal.pone.0042374 ◽

2012 ◽

Vol 7 (8) ◽

pp. e42374 ◽

Cited By ~ 40

Author(s):

Abhiram Maddi ◽

Anne Dettman ◽

Ci Fu ◽

Stephan Seiler ◽

Stephen J. Free

Keyword(s):

Cell Wall ◽

Map Kinase ◽

Neurospora Crassa ◽

Cell Wall Integrity ◽

Hyphal Fusion ◽

Map Kinase Pathway ◽

Kinase Pathway

Download Full-text

Fus3p and Kss1p Control G1 Arrest in Saccharomyces cerevisiae Through a Balance of Distinct Arrest and Proliferative Functions That Operate in Parallel With Far1p

Genetics ◽

10.1093/genetics/151.3.989 ◽

1999 ◽

Vol 151 (3) ◽

pp. 989-1004 ◽

Cited By ~ 4

Author(s):

Vera Cherkasova ◽

David M Lyons ◽

Elaine A Elion

Keyword(s):

Saccharomyces Cerevisiae ◽

Map Kinase ◽

Map Kinases ◽

G1 Arrest ◽

Multiple Functions ◽

Map Kinase Cascade ◽

Kinase Cascade ◽

Map Kinase Pathway ◽

Kinase Pathway ◽

Cyclin Genes

AbstractIn Saccharomyces cerevisiae, mating pheromones activate two MAP kinases (MAPKs), Fus3p and Kss1p, to induce G1 arrest prior to mating. Fus3p is known to promote G1 arrest by activating Far1p, which inhibits three Clnp/Cdc28p kinases. To analyze the contribution of Fus3p and Kss1p to G1 arrest that is independent of Far1p, we constructed far1 CLN strains that undergo G1 arrest from increased activation of the mating MAP kinase pathway. We find that Fus3p and Kss1p both control G1 arrest through multiple functions that operate in parallel with Far1p. Fus3p and Kss1p together promote G1 arrest by repressing transcription of G1/S cyclin genes (CLN1, CLN2, CLB5) by a mechanism that blocks their activation by Cln3p/Cdc28p kinase. In addition, Fus3p and Kss1p counteract G1 arrest through overlapping and distinct functions. Fus3p and Kss1p together increase the expression of CLN3 and PCL2 genes that promote budding, and Kss1p inhibits the MAP kinase cascade. Strikingly, Fus3p promotes proliferation by a novel function that is not linked to reduced Ste12p activity or increased levels of Cln2p/Cdc28p kinase. Genetic analysis suggests that Fus3p promotes proliferation through activation of Mcm1p transcription factor that upregulates numerous genes in G1 phase. Thus, Fus3p and Kss1p control G1 arrest through a balance of arrest functions that inhibit the Cdc28p machinery and proliferative functions that bypass this inhibition.

Download Full-text