Implications of expansin-like 3 gene in Dictyostelium morphogenesis

Kawata, Takefumi; Nakamura, Y.; Saga, Yukika; Iwade, Yumi; Ishikawa, Mariko; Sakurai, Aya; Shimada, Nao

doi:10.1186/s40064-015-0964-0

Cited by 8 publications

(6 citation statements)

References 18 publications

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“…For the tet‐off DrkA‐Myc expression construct, the ORF of the drkA gene, which harbors two introns, was amplified and inserted into pLD1ΔBX[ act15 /myc] (Kawata et al, ) to add a c‐Myc epitope tag at its C terminus. This Myc‐tagged drkA ORF region was excised from the pLD1ΔBX vector and subcloned into the pMB38 vector (Blaauw et al, ) to yield pMB38[TRE‐P min / drkA ‐myc].…”

Section: Methodsmentioning

confidence: 99%

Analysis of DrkA kinase’s role in STATa activation

et al. 2019

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Dictyostelium STATa is a homologue of metazoan signal transducers and activators of transcription (STATs) and is important for morphogenesis. STATa is activated by phosphorylation on Tyr702 when cells are exposed to extracellular cAMP. Although two tyrosine kinase-like (TKL) proteins, Pyk2 and Pyk3, have been definitively identified as STATc kinases, no kinase is known for STATa activation. Based on homology to the previously identified tyrosine-selective TKLs, we identified DrkA, a member of the TKL family and the Dictyostelium receptor-like kinase (DRK) subfamily, as a candidate STATa kinase. The drkA gene is almost exclusively expressed in prestalk A (pstA) cells, where STATa is activated. Transient over-expression of DrkA increased STATa phosphorylation, although over-expression of the protein causes a severe growth defect and cell death. Furthermore, recombinant DrkA protein is auto-phosphorylated on tyrosine and threonine residues, and an in vitro kinase assay shows that DrkA can phosphorylate STATa on Tyr702 in a STATa-SH2 (phosphotyrosine binding) domain-dependent manner. These observations strongly suggest that DrkA is one of the key regulators of STATa tyrosine phosphorylation and is consistent with it being the kinase that directly activates STATa. K E Y W O R D S cyclic AMP, Dictyostelium, phosphotyrosine signal, signal transducer and activator of transcription, tyrosine kinase-like kinase (TKL)

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Section: Methodsmentioning

confidence: 99%

Analysis of DrkA kinase’s role in STATa activation

et al. 2019

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“…Several microbial expansin proteins have been discovered and reported. These include expansin-like proteins from the Dictyostelium discoideum (Kawata et al 2015 ), BsExlx1 from Bacillus subtilis (Kerff et al 2008 ), HcExlx2 from the marine bacteria Hahella chejuensis (Lee et al 2010 ), PcExl1 from the plant pathogenic bacteria Pectobacterium carotovorum (Olarte-Lozano et al 2014 ) and ScExlx1 from the Basidiomycete fungus Schizophylum commune (Tovar-Herrera et al 2015 ). These microbial expansins have been shown to possess various capabilities which may be useful in enabling the microbes to attach and colonize plants.…”

Section: Effects Of Expansins On Specific Plant Development Stages Frmentioning

confidence: 99%

Expansins: roles in plant growth and potential applications in crop improvement

2016

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Key messageResults from various expansin related studies have demonstrated that expansins present an opportunity to improve various crops in many different aspects ranging from yield and fruit ripening to improved stress tolerance.AbstractThe recent advances in expansin studies were reviewed. Besides producing the strength that is needed by the plants, cell walls define cell shape, cell size and cell function. Expansins are cell wall proteins which consist of four sub families; α-expansin, β-expansin, expansin-like A and expansin-like B. These proteins mediate cell wall loosening and they are present in all plants and in some microbial organisms and other organisms like snails. Decades after their initial discovery in cucumber, it is now clear that these small proteins have diverse biological roles in plants. Through their ability to enable the local sliding of wall polymers by reducing adhesion between adjacent wall polysaccharides and the part they play in cell wall remodeling after cytokinesis, it is now clear that expansins are required in almost all plant physiological development aspects from germination to fruiting. This is shown by the various reports from different studies using various molecular biology approaches such as gene achieve these many roles through their non-enzymatic wall loosening ability. This paper reviews and summarizes some of the reported functions of expansins and outlines the potential uses of expansins in crop improvement programs.

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“…) and expL3 (Kawata et al . ) were amplified from the cudA null strain at similar or higher levels with compared with those from Ax2 but with reduced expression levels in the STATa null strain (Figs B, S7; Fig. S5 for ecmF , ecmH , and st15 ).…”

Section: Resultsmentioning

confidence: 98%

“…After digestion with Xba I and Not I, each DNA fragment containing the ecmF promoter fused to the lacZ coding region was gel purified and subcloned into Xba I/ Not I‐cut pLD1ΔBX‐myc (Kawata et al . ) to obtain pLD1ΔBX[ ecmF / lacZ ] and its derivatives. Alternatively, after digestion with Xba I and Xho I, each DNA fragment containing the promoter and lacZ coding region was subcloned into Xba I/ Xho I‐cut pLD1::2H3term (Shimada & Kawata ) to obtain pLD1[ ecmF / lacZ ] and its derivatives.…”

Section: Methodsmentioning

confidence: 99%

Regulation of ecmF gene expression and genetic hierarchy among STATa, CudA, and MybC on several prestalk A‐specific gene expressions in Dictyostelium

et al. 2016

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STATa, a Dictyostelium homologue of metazoan signal transducer and activator of transcription, is important for the organizer function in the tip region of the migrating Dictyostelium slug. We previously showed that ecmF gene expression depends on STATa in prestalk A (pstA) cells, where STATa is activated. Deletion and sitedirected mutagenesis analysis of the ecmF/lacZ fusion gene in wild-type and STATa null strains identified an imperfect inverted repeat sequence, ACAAATANTATTTGT, as a STATa-responsive element. An upstream sequence element was required for efficient expression in the rear region of pstA zone; an element downstream of the inverted repeat was necessary for sufficient prestalk expression during culmination. Band shift analyses using purified STATa protein detected no sequence-specific binding to those ecmF elements. The only verified upregulated target gene of STATa is cudA gene; CudA directly activates expL7 gene expression in prestalk cells. However, ecmF gene expression was almost unaffected in a cudA null mutant. Several previously reported putative STATa target genes were also expressed in cudA null mutant but were downregulated in STATa null mutant. Moreover, mybC, which encodes another transcription factor, belonged to this category, and ecmF expression was downregulated in a mybC null mutant. These findings demonstrate the existence of a genetic hierarchy for pstA-specific genes, which can be classified into two distinct STATa downstream pathways, CudA dependent and independent. The ecmF expression is indirectly upregulated by STATa in a CudAindependent activation manner but dependent on MybC, whose expression is positively regulated by STATa.

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Implications of expansin-like 3 gene in Dictyostelium morphogenesis

Cited by 8 publications

References 18 publications

Analysis of DrkA kinase’s role in STATa activation

Analysis of DrkA kinase’s role in STATa activation

Expansins: roles in plant growth and potential applications in crop improvement

Regulation of ecmF gene expression and genetic hierarchy among STATa, CudA, and MybC on several prestalk A‐specific gene expressions in Dictyostelium

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