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1.
Genes (Basel) ; 10(5)2019 05 01.
Article in English | MEDLINE | ID: mdl-31052454

ABSTRACT

Identification and characterization of protein complexes and interactomes has been essential to the understanding of fundamental nuclear processes including transcription, replication, recombination, and maintenance of genome stability. Despite significant progress in elucidation of nuclear proteomes and interactomes of organisms such as yeast and mammalian systems, progress in other models has lagged. Protists, including the alveolate ciliate protozoa with Tetrahymena thermophila as one of the most studied members of this group, have a unique nuclear biology, and nuclear dimorphism, with structurally and functionally distinct nuclei in a common cytoplasm. These features have been important in providing important insights about numerous fundamental nuclear processes. Here, we review the proteomic approaches that were historically used as well as those currently employed to take advantage of the unique biology of the ciliates, focusing on Tetrahymena, to address important questions and better understand nuclear processes including chromatin biology of eukaryotes.


Subject(s)
Ciliophora Infections/genetics , Nuclear Proteins/genetics , Proteomics , Tetrahymena thermophila/genetics , Cell Nucleus/genetics , Cell Nucleus/parasitology , Chromatin/genetics , Chromatin/parasitology , Ciliophora Infections/parasitology , Cytoplasm/genetics , Cytoplasm/parasitology , Humans , Tetrahymena thermophila/pathogenicity
2.
J Eukaryot Microbiol ; 62(5): 605-13, 2015.
Article in English | MEDLINE | ID: mdl-25733238

ABSTRACT

The activities of Tetrahymena corlissi, Tetrahymena thermophila, and Tetrahymena canadensis were studied in coculture with cell lines of insects, fish, amphibians, and mammals. These ciliates remained viable regardless of the animal cell line partner. All three species could engulf animal cells in suspension. However, if the animal cells were monolayer cultures, the monolayers were obliterated by T. corlissi and T. thermophila. Both fibroblast and epithelial monolayers were destroyed but the destruction of human cell monolayers was done more effectively by T. thermophila. By contrast, T. canadensis was unable to destroy any monolayer. At 4 °C T. thermophila and T. corlissi did not carryout phagocytosis and did not destroy monolayers, whereas T. canadensis was able to carryout phagocytosis but still could not destroy monolayers. Therefore, monolayer destruction appeared to require phagocytosis, but by itself this was insufficient. In addition, the ciliates expressed a unique swimming behavior. Tetrahymena corlissi and T. thermophila swam vigorously and repeatedly into the monolayer, which seemed to loosen or dislodge cells, whereas T. canadensis swam above the monolayer. Therefore, differences in swimming behavior might explain why T. corlissi has been reported to be a pathogen but T. canadensis has not.


Subject(s)
Tetrahymena/physiology , Tetrahymena/pathogenicity , Amphibians/parasitology , Animals , Axenic Culture , Cell Culture Techniques , Cell Line , Fishes/parasitology , HeLa Cells , Humans , Insecta/cytology , Insecta/parasitology , Mammals/parasitology , Phagocytosis , Swimming , Temperature , Tetrahymena/classification , Tetrahymena thermophila/pathogenicity , Tetrahymena thermophila/physiology , Tetrahymena thermophila/ultrastructure
3.
Eukaryot Cell ; 12(8): 1080-6, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23729382

ABSTRACT

ADF/cofilin is a highly conserved actin-modulating protein. Reorganization of the actin cytoskeleton in vivo through severing and depolymerizing of F-actin by this protein is essential for various cellular events, such as endocytosis, phagocytosis, cytokinesis, and cell migration. We show that in the ciliate Tetrahymena thermophila, the ADF/cofilin homologue Adf73p associates with actin on nascent food vacuoles. Overexpression of Adf73p disrupted the proper localization of actin and inhibited the formation of food vacuoles. In vitro, recombinant Adf73p promoted the depolymerization of filaments made of T. thermophila actin (Act1p). Knockout cells lacking the ADF73 gene are viable but grow extremely slowly and have a severely decreased rate of food vacuole formation. Knockout cells have abnormal aggregates of actin in the cytoplasm. Surprisingly, unlike the case in animals and yeasts, in Tetrahymena, ADF/cofilin is not required for cytokinesis. Thus, the Tetrahymena model shows promise for future studies of the role of ADF/cofilin in vivo.


Subject(s)
Actins/metabolism , Cofilin 1/genetics , Microfilament Proteins/genetics , Phagocytosis/genetics , Tetrahymena thermophila/growth & development , Tetrahymena thermophila/metabolism , Actin Cytoskeleton/metabolism , Ciliophora Infections/genetics , Ciliophora Infections/microbiology , Cofilin 1/metabolism , Cytokinesis/genetics , Gene Knockout Techniques , Sequence Homology, Amino Acid , Tetrahymena thermophila/pathogenicity , Vacuoles/metabolism
4.
J Evol Biol ; 24(12): 2563-73, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21902750

ABSTRACT

Temporal resource fluctuations could affect the strength of antagonistic coevolution through population dynamics and costs of adaptation. We studied this by coevolving the prey bacterium Serratia marcescens with the predatory protozoa Tetrahymena thermophila in constant and pulsed-resource environments for approximately 1300 prey generations. Consistent with arms race theory, the prey evolved to be more defended, whereas the predator evolved to be more efficient in consuming the bacteria. Coevolutionary adaptations were costly in terms of reduced prey growth in resource-limited conditions and less efficient predator growth on nonliving resource medium. However, no differences in mean coevolutionary changes or adaptive costs were observed between environments, even though resource pulses increased fluctuations and mean densities of coevolving predator populations. Interestingly, a surface-associated prey defence mechanism (bacterial biofilm), to which predators were probably unable to counter-adapt, evolved to be stronger in pulsed-resource environment. These results suggest that temporal resource fluctuations can increase the asymmetry of antagonistic coevolution by imposing stronger selection on one of the interacting species.


Subject(s)
Biological Evolution , Serratia marcescens/growth & development , Tetrahymena thermophila/pathogenicity , Adaptation, Physiological , Biofilms , Culture Media , Environment , Microbiological Techniques/methods , Serratia marcescens/physiology , Species Specificity , Tetrahymena thermophila/growth & development , Tetrahymena thermophila/physiology , Time Factors
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