Screening of binding proteins that interact with two components of the HOG MAPK pathway by the yeast two-hybrid method in Colletotrichum siamense

Liao, Xiaomiao; Long, Xiping; He, Qiang; Miao, Song; Xiao, Li; Liu, Wenbo; Lin, Chunhua; Miao, Weiguo

doi:10.1007/s10658-021-02205-y

Cited by 4 publications

(5 citation statements)

References 22 publications

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“…During the identification process of AwSakA, we found that SakA is evolutionarily conserved in the Aspergillus, and a comparative sequence analysis of all fungal species showed that SakA is highly similar and that it shares the same highly conserved protein kinase structural domain. This is consistent with the study showing that SakA encodes part of the MAPK gene family in the fungi A. alternata [16] and P. verrucosum [18]. We identified the gene function of AwSakA and investigated the growth and conidial and OTA production of ∆AwSakA.…”

Section: Discussionsupporting

confidence: 90%

“…In fungal cells, the MAPK signaling pathway plays a key role in regulating development by transmitting extracellular signals to the nucleus after sensing stimuli from the surrounding environment [15]. The MAPK pathway is commonly associated with oxidative responses, osmoregulation and aspects of fungicide susceptibility in plant pathogens [16][17][18]. MAPKs, also known as SAPKs (stress-activated protein kinases), generate multiple stress responses through environmental signaling.…”

Section: Introductionmentioning

confidence: 99%

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SakA Regulates Morphological Development, Ochratoxin A Biosynthesis and Pathogenicity of Aspergillus westerdijkiae and the Response to Different Environmental Stresses

Wang

et al. 2023

Toxins

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Ochratoxin A (OTA), as a common mycotoxin, has seriously harmful effects on agricultural products, livestock and humans. There are reports on the regulation of SakA in the MAPK pathway, which regulates the production of mycotoxins. However, the role of SakA in the regulation of Aspergillus westerdijkiae and OTA production is not clear. In this study, a SakA deletion mutant (ΔAwSakA) was constructed. The effects of different concentrations of D-sorbitol, NaCl, Congo red and H2O2 on the mycelia growth, conidia production and biosynthesis of OTA were investigated in A. westerdijkiae WT and ΔAwSakA. The results showed that 100 g/L NaCl and 3.6 M D-sorbitol significantly inhibited mycelium growth and that a concentration of 0.1% Congo red was sufficient to inhibit the mycelium growth. A reduction in mycelium development was observed in ΔAwSakA, especially in high concentrations of osmotic stress. A lack of AwSakA dramatically reduced OTA production by downregulating the expression of the biosynthetic genes otaA, otaY, otaB and otaD. However, otaC and the transcription factor otaR1 were slightly upregulated by 80 g/L NaCl and 2.4 M D-sorbitol, whereas they were downregulated by 0.1% Congo red and 2 mM H2O2. Furthermore, ΔAwSakA showed degenerative infection ability toward pears and grapes. These results suggest that AwSakA is involved in the regulation of fungal growth, OTA biosynthesis and the pathogenicity of A. westerdijkiae and could be influenced by specific environmental stresses.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

SakA Regulates Morphological Development, Ochratoxin A Biosynthesis and Pathogenicity of Aspergillus westerdijkiae and the Response to Different Environmental Stresses

Wang

et al. 2023

Toxins

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“…Our previous study showed that a fatty acid hydroxylase is the CsPbs2-interacting protein by the Y2H system and pull-down method ( 22 ). To characterize the function of this gene, the whole coding sequence was obtained from the C. siamense HN08 strain by PCR and RT-PCR.…”

Section: Resultsmentioning

confidence: 99%

“…The sequence of CsSCS7 was obtained previously from the yeast cDNA library of C. siamense HN08 using the CsPbs2 protein of C. siamense as the bait protein ( 22 ). The CsSCS7 coding regions and upstream and downstream sequences were obtained from the HN08 genome database and transcriptome database through a local BLAST search.…”

Section: Methodsmentioning

confidence: 99%

“…C. siamense is reportedly one of the major pathogenic species of many tropical or subtropical crops, including rubber trees ( 21 ). Our research group previously screened and identified the fatty acid hydroxylase CsSCS7, which exhibit 51% identity to S. cerevisiae SCS7, as the interacting protein of the core member CsPbs2 of the HOG MAPK in C. siamense ( 22 ). In this study, the function of CsSCS7 in C. siamense was characterized.…”

Section: Introductionmentioning

confidence: 99%

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The fatty acid 2-hydroxylase CsSCS7 is a key hyphal growth factor and potential control target in Colletotrichum siamense

Xi,

Long,

Song

et al. 2024

mBio

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SCS7 is a fatty acid 2-hydroxylase required for the synthesis of inositol phosphorylceramide but is not essential for normal growth in Saccharomyces cerevisiae . Here, we demonstrate that the Colletotrichum siamense SCS7 homolog CsSCS7 plays a key role in hyphal growth. The CsSCS7 deletion mutant showed strong hyphal growth inhibition, small conidia, and marginally reduced sporulation and also resulted in a sharp reduction in the full virulence and increasing the fungicide sensitivity. The three protein domains (a cytochrome b5 domain, a transmembrane domain, and a hydroxylase domain) are important to CsSCS7 protein function in hyphal growth. The fatty acid assay results revealed that the CsSCS7 gene is important for balancing the contents of multiple mid-long- and short-chain fatty acids. Additionally, the retarded growth and virulence of C. siamense Δ CsSCS7 can be recovered partly by the reintroduction of homologous sequences from Magnaporthe oryzae and Fusarium graminearum but not SCS7 of S. cerevisiae . In addition, the spraying of C. siamense with naked CsSCS7 -double-stranded RNA (dsRNAs), which leads to RNAi, increases the inhibition of hyphal growth and slightly decreases disease lesions. Then, we used nano material Mg-Al-layered double hydroxide as carriers to deliver dsRNA, which significantly enhanced the control effect of dsRNA, and the lesion area was obviously reduced. These data indicated that CsSCS7 is an important factor for hyphal growth and affects virulence and may be a potential control target in C. siamense and even in filamentous plant pathogenic fungi. IMPORTANCE CsSCS7, which is homologous to yeast fatty acid 2-hydroxylase SCS7, was confirmed to play a key role in the hyphal growth of Colletotrichum siamense and affect its virulence. The CsSCS7 gene is involved in the synthesis and metabolism of fatty acids. Homologs from the filamentous fungi Magnaporthe oryzae and Fusarium graminearum can recover the retarded growth and virulence of C. siamense ΔCsSCS7. The spraying of double-stranded RNAs targeting CsSCS7 can inhibit hyphal growth and reduce the disease lesion area to some extent. After using nano material Mg-Al layered double hydroxide as carrier, the inhibition rates were significantly increased. We demonstrated that CsSCS7 is an important factor for hyphal growth and affects virulence and may be a potential control target in C. siamense and even in filamentous plant pathogenic fungi.

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Screening Proteins That Interact With AcHog1 and the Functional Analysis of AcSko1 in Aspergillus cristatus

Shao,

Liu,

Liu

et al. 2024

Journal of Basic Microbiology

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Aspergillus cristatus is a dominant fungus formed during the “flowering” process of Fuzhuan brick tea. Previous research has established that the sporulation of Aspergillus nidulans, a model organism of filamentous fungi, is regulated by light. However, the sporulation of A. cristatus is dependent on osmotic stress. In a previous study, we used pull‐down and mass spectrometry to identify proteins that interacted with AcHog1 in A. cristatus when cultured under different conditions of osmotic stress. In the present study, we analyzed the proteins we identified previously to investigate their functional role. The AA1E3BER4 protein was located downstream of Hog1 in the HOG branch pathway and was identified that was regulated by AcHog1. Furthermore, yeast two‐hybrid analysis showed that AA1E3BER4 interacted with AcHog1. In addition, we knocked out and complemented the Acsko1 gene encoding the AA1E3BER4 protein. We found that the number of sexual and asexual spores were downregulated by 3.81‐ and 4.57‐fold, respectively, in the ΔAcsko1 strain. The sensitivity of the ΔAcsko1 strain to sorbitol and sucrose, as regulators of osmotic stress, increased, and the sensitivity to high sucrose was higher than that of sorbitol. Acsko1 also regulated the response of A. cristatus to oxidative stress, Congo red, and SDS (sodium dodecyl sulfate). In addition, the deletion of Acsko1 significantly increased the pigment of the ΔAcsko1 strain. This is the first study to report the role of the sko1 gene in oxidative stress, stress‐induced damage to the cell wall, and pigment in Aspergillus cristatus.

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Screening of binding proteins that interact with two components of the HOG MAPK pathway by the yeast two-hybrid method in Colletotrichum siamense

Cited by 4 publications

References 22 publications

SakA Regulates Morphological Development, Ochratoxin A Biosynthesis and Pathogenicity of Aspergillus westerdijkiae and the Response to Different Environmental Stresses

SakA Regulates Morphological Development, Ochratoxin A Biosynthesis and Pathogenicity of Aspergillus westerdijkiae and the Response to Different Environmental Stresses

The fatty acid 2-hydroxylase CsSCS7 is a key hyphal growth factor and potential control target in Colletotrichum siamense

Screening Proteins That Interact With AcHog1 and the Functional Analysis of AcSko1 in Aspergillus cristatus

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