Ras small GTPases act as molecular switches in various cellular signaling pathways, including cell migration, proliferation, and differentiation. Three Rap proteins are present in Dictyostelium; RapA, RapB, and RapC. RapA and RapC have been reported to have opposing functions in the control of cell adhesion and migration. Here, we investigated the role of RapB, a member of the Ras GTPase subfamily in Dictyostelium, focusing on its involvement in cell adhesion, migration, and developmental processes. This study revealed that RapB, similar to RapA, played a crucial role in regulating cell morphology, adhesion, and migration. rapB null cells, which were generated by CRISPR/Cas9 gene editing, displayed altered cell size, reduced cell-substrate adhesion, and increased migration speed during chemotaxis. These phenotypes of rapB null cells were restored by the expression of RapB and RapA, but not RapC. Consistent with these results, RapB, similar to RapA, failed to rescue the phenotypes of rapC null cells, spread morphology, increased cell adhesion, and decreased migration speed during chemotaxis. Multicellular development of rapB null cells remained unaffected. These results suggest that RapB is involved in controlling cell morphology and cell adhesion. Importantly, RapB appears to play an inhibitory role in regulating the migration speed during chemotaxis, possibly by controlling cell-substrate adhesion, resembling the functions of RapA. These findings contribute to the understanding of the functional relationships among Ras subfamily proteins.
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Intracellular Calcium Responses to External Calcium Stimuli in Dictyostelium Dahyeon Kim, Jiseong Seo, Taeck Joong Jeon Journal of Microbiology and Biotechnology.2025;[Epub] CrossRef
Three novel, Gram-stain-positive, obligate aerobic, catalase- and oxidase-positive bacterial strains, designated B2O-1(T), T2O-4(T), and 0.2-SM1T-5(T), were isolated from jeotgal, a traditional Korean fermented seafood. Strains B2O-1(T), T2O-4(T), and 0.2-SM1T-5(T) exhibited distinct colony colors, characterized by pink, yellow, and red opaque circular colonies, respectively. Phylogenetic analysis revealed that three strains formed a paraphyletic clade within the genus Sporosarcina and shared < 99.0% similarity with Sporosarcina aquimarina KCTC 3840(T) and Sporosarcina saromensis KCTC 13119(T) in their 16S rRNA gene sequences. The three strains exhibiting Orthologous Average Nucleotide Identity values < 79.3% and digital DNA-DNA hybridization values < 23.1% within the genus Sporosarcina affirmed their distinctiveness. Strains B2O-1(T), T2O-4(T), and 0.2-SM1T-5(T) contained MK-7 as a sole respiratory menaquinone and A4α type peptidoglycan based on lysine with alanine, glutamic acid, and aspartic acid. The common polar lipids include diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine.
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Aconitum carmichaeli Debx. (Ranunculaceae) is a potential
source of an important herbal drug named “Fuzi”, which is
derived from the lateral root of the plant. Increased therapeutic
usage resulted in the great demand for artificial cultivation
of A. carmichaeli, however, the obstacles caused by
continuous cropping is a serious problem. Continuous cropping
has shown to affect the soil biological and non-biological
factors. The current study attempted to discover the variations
of microbial communities and soil properties in shortterm
continuous cropping of A. carmichaeli. An experimental
procedure with A. carmichaeli planted two years continuously
was established. The variation of the soil microbial community,
disease incidence, soil properties, and the correlation
between soil microbe and disease incidence were investigated.
The disease incidence increased during the continuous cropping
of A. carmichaeli. The PCoA and LefSe results indicated
that fungal communities in rhizosphere soil were altered during
the short-term continuous croppingand the bacterial community
was disturbed by the cultivation of A. carmichaeli,
however, in the following two years of continuous cropping
period, the soil bacterial community has not changed obviously.
Proportions of some fungal and bacterial genera were
varied significantly (p < 0.05), and some genera of microflora
showed a significant correlation with adisease incidence of
A. carmichaeli. Microorganisms contributing to community
composition discrepancy were also elucidated. Continuous
cropping of A. carmichaeli disturbed the rhizosphere soil microbial
community and altered the soil chemical parameters
and soil pH. These variations in soil may be related to the
occurrence of plant diseases. The current study will not only
provide theoretical and experimental evidence for the A.
carmichaeli continuous cropping obstacles but will also contribute
to A. carmichaeli agricultural production and soil
improvement.
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FERM domain-containing proteins are involved in diverse
biological and pathological processes, including cell-substrate
adhesion, cell-cell adhesion, multicellular development,
and cancer metastasis. In this study, we determined the functions
of FrmB, a FERM domain-containing protein, in the
cell morphology, cell adhesion, and multicellular development
of Dictyostelium cells. Our results show that FrmB appears
to play an important role in regulating the size of developmental
structures. frmB null cells showed prolonged aggregation
during development, resulting in increased size of developmental
structures, such as mounds and fruiting bodies,
compared to those of wild-type cells, whereas FrmB overexpressing
cells exhibited decreased size of developmental
structures. These results suggest that FrmB may be necessary
for limiting the sizes of developmental structures. Loss of
FrmB also resulted in decreased cell-substrate adhesion and
slightly increased cell area, suggesting that FrmB had important
roles in the regulation of cell adhesion and cell morphology.
These studies would contribute to our understanding
of the intertwined and overlapped functions of FERM
domain-containing proteins.
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Establishment of cell polarity is mediated by a series of signal-ing molecules that are asymmetrically activated or localized in the cell upon extracellular stimulation. To understand the mechanism that mediates anterior/posterior asymmetric localization of RapGAP3 during migration, we determined the minimally required amino acids in the I/LWEQ domain that cause posterior localization and found that the minimal region of the F-actin binding domain for posterior localiza-tion could, with some additional deletion at the C-terminal, localize to the anterior. Analysis of the localization and trans-location kinetics to the cell cortex of the truncated proteins suggests that the required regions for anterior/posterior lo-calization might have a preferential binding affinity to pre- existing F-actins at the rear and lateral sides of the cell or newly formed F-actins at the front of the cell, leading to dis-tinct differential sites of the cell.
Cell movement involves a coordinated regulation of the cytoskeleton, F-actin-mediated protrusions at the front and myosin-mediated contraction of the posterior of a cell. The small GTPase Rap1 functions as a key regulator in the spatial and temporal control of cytoskeleton reorganization for cell migration. This review outlines the establishment of cell polarity by differential localizations of the cytoskeleton and discusses the spatial and temporal regulation of cytoskeleton
reorganization via the Rap1 signaling pathway during chemotaxis with a focus on recent advances in the study of chemotaxis using a simple eukaryotic model organism, Dictyostelium discoideum.
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