scholarly journals Hyphal compartmentalization and sporulation in Streptomyces require the conserved cell division protein SepX

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Matthew J. Bush ◽  
Kelley A. Gallagher ◽  
Govind Chandra ◽  
Kim C. Findlay ◽  
Susan Schlimpert
Keyword(s):  
Life Cycle ◽  
Cell Division ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Important Determinant ◽  
Specific Protein ◽  
Cross Wall ◽  
Developmental Life Cycle ◽  
Cell Division Protein ◽  
Cellular Development

AbstractFilamentous actinobacteria such as Streptomyces undergo two distinct modes of cell division, leading to partitioning of growing hyphae into multicellular compartments via cross-walls, and to septation and release of unicellular spores. Specific determinants for cross-wall formation and the importance of hyphal compartmentalization for Streptomyces development are largely unknown. Here we show that SepX, an actinobacterial-specific protein, is crucial for both cell division modes in Streptomyces venezuelae. Importantly, we find that sepX-deficient mutants grow without cross-walls and that this substantially impairs the fitness of colonies and the coordinated progression through the developmental life cycle. Protein interaction studies and live-cell imaging suggest that SepX contributes to the stabilization of the divisome, a mechanism that also requires the dynamin-like protein DynB. Thus, our work identifies an important determinant for cell division in Streptomyces that is required for cellular development and sporulation.

scholarly journals Multicellular growth and sporulation in filamentous actinobacteria require the conserved cell division protein SepX

2021 ◽  
Author(s):  
Susan Schlimpert ◽  
Matthew James Bush ◽  
Kelley Ann Gallagher ◽  
Govind Chandra ◽  
Kim Findlay
Keyword(s):  
Cell Division ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Important Determinant ◽  
Specific Protein ◽  
Cross Wall ◽  
Specific Cell ◽  
Multicellular Development ◽  
Developmental Life Cycle ◽  
Cell Division Protein

Filamentous actinobacteria like Streptomyces undergo two distinct modes of cell division, leading to the partitioning of growing hyphae into multicellular compartments via cross-walls and to the septation and release of unicellular spores. While some progress has been made towards the regulation of sporulation-specific cell division, specific determinants for cross-wall formation and the importance of hyphal compartmentalization for Streptomyces development have remained unknown. Here we describe SepX, an actinobacterial-specific protein that is crucial for both cell division events in Streptomyces. We show that sepX-deficient mutants grow without cross-walls and that this substantially impairs the fitness of colonies and the coordinated progression through the developmental life cycle. Protein interaction studies and live-cell imaging suggest that SepX functions to spatially stabilize the divisome, a mechanism that also requires the dynamin-like protein DynB. Collectively, this work identifies an important determinant for cell division in filamentous actinobacteria that is required for multicellular development and sporulation.

scholarly journals Live cell imaging of the HIV-1 life cycle

2008 ◽  
Vol 16 (12) ◽  
pp. 580-587 ◽  
Author(s):  
Edward M. Campbell ◽  
Thomas J. Hope
Keyword(s):  
Life Cycle ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Hiv 1

Live Cell Imaging of the Schizosaccharomyces pombe Sexual Life Cycle

2017 ◽  
Vol 2017 (10) ◽  
pp. pdb.prot090225 ◽  
Author(s):  
Laura Merlini ◽  
Aleksandar Vjestica ◽  
Omaya Dudin ◽  
Felipe Bendezú ◽  
Sophie G. Martin
Keyword(s):  
Life Cycle ◽  
Schizosaccharomyces Pombe ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Sexual Life ◽  
Sexual Life Cycle

scholarly journals Live cell imaging of the hyperthermophilic archaeon Sulfolobus acidocaldarius identifies complementary roles for two ESCRTIII homologues in ensuring a robust and symmetric cell division

2020 ◽  
Author(s):  
Andre Arashiro Pulschen ◽  
Delyan R. Mutavchiev ◽  
Kim Nadine Sebastian ◽  
Jacques Roubinet ◽  
Marc Roubinet ◽  
...  
Keyword(s):  
Cell Division ◽  
Cell Biology ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Halophilic Archaea ◽  
Live Cell ◽  
Tight Coupling ◽  
Dna Compaction ◽  
Cellular Processes ◽  
Daughter Cells

Live-cell imaging has revolutionized our understanding of dynamic cellular processes in bacteria and eukaryotes. While similar techniques have recently been applied to the study of halophilic archaea, our ability to explore the cell biology of thermophilic archaea is limited, due to the technical challenges of imaging at high temperatures. Here, we report the construction of the Sulfoscope, a heated chamber that enables live-cell imaging on an inverted fluorescent microscope. Using this system combined with thermostable fluorescent probes, we were able to image Sulfolobus cells as they divide, revealing a tight coupling between changes in DNA compaction, segregation and cytokinesis. By imaging deletion mutants, we observe important differences in the function of the two ESCRTIII proteins recently implicated in cytokinesis. The loss of CdvB1 compromises cell division, causing occasional division failures and fusion of the two daughter cells, whereas the deletion of cdvB2 leads to a profound loss of division symmetry, generating daughter cells that vary widely in size and eventually generating ghost cells. These data indicate that DNA separation and cytokinesis are coordinated in Sulfolobus, as is the case in eukaryotes, and that two contractile ESCRTIII polymers perform distinct roles to ensure that Sulfolobus cells undergo a robust and symmetrical division. Taken together, the Sulfoscope has shown to provide a controlled high temperature environment, in which cell biology of Sulfolobus can be studied in unprecedent details.

scholarly journals Kinesin-8B controls basal body function and flagellum formation and is key to malaria parasite transmission

2019 ◽  
Author(s):  
Mohammad Zeeshan ◽  
David J. P. Ferguson ◽  
Steven Abel ◽  
Alana Burrrell ◽  
Edward Rea ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Life Cycle ◽  
Live Cell Imaging ◽  
Malaria Parasite ◽  
Cell Imaging ◽  
Male Gamete ◽  
Live Cell ◽  
Basal Bodies ◽  
Parasite Transmission ◽  
Parasite Life Cycle

AbstractEukaryotic flagella are conserved microtubule-based organelles that drive cell motility. Plasmodium, the causative agent of malaria, has a single flagellate stage: the male gamete in the mosquito. Three rounds of endomitotic division together with an unusual mode of flagellum assembly rapidly produce eight motile gametes. These processes are tightly coordinated but their regulation is poorly understood. To understand this important developmental stage, we studied the function and location of the microtubule-based motor kinesin-8B, using gene-targeting, electron microscopy and live cell imaging. Deletion of the kinesin-8B gene showed no effect on mitosis but disrupted 9+2 axoneme assembly and flagellum formation during male gamete development and also completely ablated parasite transmission. Live cell imaging showed that kinesin-8B-GFP did not colocalise with kinetochores in the nucleus but instead revealed dynamic, cytoplasmic localisation with the basal bodies and the assembling axoneme during flagellum formation. We thus uncovered an unexpected role for kinesin-8B in parasite flagellum formation that is vital for the parasite life cycle.

scholarly journals Microtubule Probe for Correlative Super-Resolution Fluorescence and Electron Microscopy

2018 ◽  
Author(s):  
Xiaohe Tian ◽  
Cesare De Pace ◽  
Lorena Ruiz-Perez ◽  
Bo Chen ◽  
Rina Su ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Division ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Synchronous Fluorescence ◽  
Live Cell ◽  
Fluorescence And Electron Microscopy ◽  
20 Nm ◽  
In Cells

We report a versatile cyclometalated Iridium (III) complex probe that achieves synchronous fluorescence-electron microscopy correlation to reveal microtubule ultrastructure in cells. The selective insertion of probe between repeated α and β units of microtubule triggers remarkable fluorescent enhancement, and high TEM contrast due to the presence of heavy Ir ions. The highly photostable probe allows live cell imaging of tubulin localization and motion during cell division with an resolution of 20 nm, and under TEM imaging reveals the αβ unit interspace of 45Å of microtubule in cells.

scholarly journals Revealing spatio-temporal dynamics with long-term trypanosomatid live-cell imaging

2021 ◽  
Author(s):  
Richard S Muniz ◽  
Paul C Campbell ◽  
Thomas E Sladewski ◽  
Lars D Renner ◽  
Christopher L de Graffenried
Keyword(s):  
Cell Division ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Temporal Dynamics ◽  
Live Cell ◽  
Three Dimensions ◽  
Imaging Method ◽  
Differential Rate ◽  
Pdms Stamp

Trypanosoma brucei, the causative agent of human African trypanosomiasis, employs a flagellum for dissemination within the parasite's mammalian and insect hosts. T. brucei cells are highly motile in culture and must be able to move in all three dimensions for reliable cell division. These characteristics have made long-term microscopic imaging of live T. brucei cells challenging, which has limited our understanding of a variety of important cell-cycle events. To address this issue, we have devised an imaging approach that confines cells to small volumes that can be imaged continuously for up to 24 h. This system employs cast agarose microwells generated using a PDMS stamp that can be made with different dimensions to maximize cell viability and imaging quality. Using this approach, we have imaged individual T. brucei through multiple rounds of cell division with high spatial and temporal resolution. We have employed this method to study the differential rate of T. brucei daughter cell division and show that the approach is compatible with loss-of-function experiments such as small molecule inhibition and RNAi. We have also developed a strategy that employs in-well "sentinel" cells to monitor potential toxicity due to imaging. This live-cell imaging method will provide a novel avenue for studying a wide variety of cellular events in trypanosomatids that have previously been inaccessible.

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