Regulation of Macronuclear DNA Content in <i>Tetrahymena thermophila</i>*†

Doerder, F. Paul

doi:10.1111/j.1550-7408.1979.tb02726.x

Cited by 53 publications

(4 citation statements)

References 5 publications

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“…Tetrahymena's amitotic (AM) MAC is characterized by a G1-S-G2-AM cell division cycle, which roughly coincides with the regular G1-S-G2-M pattern seen in mitosis (Figure 6E; Flickinger, 1965;Woodard et al, 1972;Doerder, 1979;Cole and Sugai, 2012). The MIC undergoes more conventional mitosis, though with no apparent G1 interval.…”

Section: Discussionmentioning

confidence: 59%

“…The MIC undergoes more conventional mitosis, though with no apparent G1 interval. MIC S phase and mitosis are temporally out of phase with the MAC nuclear cycle (Figure 6E; Flickinger, 1965;Woodard et al, 1972;Doerder, 1979). The highly synchronized cell populations we can obtain by CCE will allow us to address important questions in Tetrahymena biology.…”

Section: Discussionmentioning

confidence: 93%

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An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

Liu

Nan

Niu

et al. 2021

Front. Cell Dev. Biol.

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Cell synchronization is a powerful tool to understand cell cycle events and its regulatory mechanisms. Counter-flow centrifugal elutriation (CCE) is a more generally desirable method to synchronize cells because it does not significantly alter cell behavior and/or cell cycle progression, however, adjusting specific parameters in a cell type/equipment-dependent manner can be challenging. In this paper, we used the unicellular eukaryotic model organism, Tetrahymena thermophila as a testing system for optimizing CCE workflow. Firstly, flow cytometry conditions were identified that reduced nuclei adhesion and improved the assessment of cell cycle stage. We then systematically examined how to achieve the optimal conditions for three critical factors affecting the outcome of CCE, including loading flow rate, collection flow rate and collection volume. Using our optimized workflow, we obtained a large population of highly synchronous G1-phase Tetrahymena as measured by 5-ethynyl-2′-deoxyuridine (EdU) incorporation into nascent DNA strands, bulk DNA content changes by flow cytometry, and cell cycle progression by light microscopy. This detailed protocol can be easily adapted to synchronize other eukaryotic cells.

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Section: Discussionmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 93%

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

Liu

Nan

Niu

et al. 2021

Front. Cell Dev. Biol.

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“…Analysis of variances of DNA content in pre‐replication vs. post‐replication cells showed that in Paramecium the coefficient of variation is halved during the replication process (Berger and Schmidt 1978). In contrast, macronuclear DNA content in Tetrahymena is doubled and the coefficient of variation remains constant (Doerder 1979). We postulated that all Paramecium cells were making the same amount of DNA.…”

Section: Regulation Of Macronuclear Dna Contentmentioning

confidence: 99%

“…This system for regulation of DNA content is very different from that in Tetrahymena in which the prereplication DNA content (i.e. genome copy number or gene dosage) determines whether 0, 1 or 2 complete rounds of DNA replication occur within the cell cycle (Cleffmann 1968, 1975; Cleffmann, Reuther, and Seyfert 1979; Doerder 1979; Doerder and DeBault 1978).…”

Section: Regulation Of Macronuclear Dna Contentmentioning

confidence: 99%

Riding The Ciliate Cell Cycle—A Thirty‐Five‐Year Prospective^a

Berger

2001

J Eukaryotic Microbiology

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Studies of the ciliate cell cycle have moved from early examination of its biochemistry with heat-synchronized Tetrahymena through descriptive studies of Paramecium using small synchronous cell samples. These studies described what happens during the cell cycle and provided some initial insights into control, especially the idea that there was a point at which cells became committed to division. This early work was followed by an analytical phase in which the same small sample techniques, combined with gene mutations, were used to tease apart some major features of the regulation of cell growth kinetics, including regulation of macronuclear DNA content and regulation of cell size, the control of timing of initiation of macronuclear DNA synthesis, and the control of commitment to division in Paramecium. The availability of new molecular genetic approaches and new means of manipulating cells en masse made it possible to map out some of the basic features of the molecular biology of cell cycle regulation in ciliates. The challenge before us is to move beyond the 'me-too-ism' of validating the presence of basic molecular regulative machinery underlying the cell cycle in ciliates to a deeper analysis of the role of specific molecules in processes unique to ciliates or to analysis of the role of regulatory molecules in the control of cell process that can be uniquely well studied in ciliates.

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Tetrahymena

Coyne¹

2011

Encyclopedia of Life Sciences

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Tetrahymena is a genus of mostly free‐living ciliated protozoa that is intensively employed to investigate and solve fundamental problems in molecular, cellular and developmental biology. Like all ciliates, Tetrahymena contains separate germline and somatic nuclei, known as the micronucleus and macronucleus, respectively. The macronucleus is derived from a copy of the micronucleus through a process that involves extensive programmed whole‐genome rearrangement and is under intensive study. The most highly developed experimental model species is Tetrahymena thermophila , which can be readily manipulated using the tools of genetics, molecular biology, cell biology and biochemistry. Notable discoveries made using Tetrahymena include the structure of telomeres and telomerase, self‐splicing RNA, the first microtubular motor and the link between histone acetylation and gene regulation. The approximately 104 Megabase macronuclear genome of T. thermophila has been sequenced and annotated; the micronuclear genome sequence will be completed soon. Key Concepts: Tetrahymena are large, elaborate eukaryotic cells with many experimental advantages and well‐suited to the study of cellular structure, division and development. Tetrahymena have been studied for nearly a century and been the source of a number of groundbreaking discoveries. A characteristic feature of Tetrahymena , and the basis of much biological interest, is the separation of germline and somatic genetic functions into separate nuclei. Programmed genome rearrangement in Tetrahymena shares common mechanistic features with heterochromatic gene silencing in other eukaryotes. Applying the modern tools of genomics and proteomics has facilitated research with Tetrahymena and opened up new areas of investigation.

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Regulation of Macronuclear DNA Content in *Tetrahymena thermophila**†

Cited by 53 publications

References 5 publications

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

Riding The Ciliate Cell Cycle—A Thirty‐Five‐Year Prospective^a

Tetrahymena

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Regulation of Macronuclear DNA Content in Tetrahymena thermophila*†

Cited by 53 publications

References 5 publications

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells

Riding The Ciliate Cell Cycle—A Thirty‐Five‐Year Prospectivea

Tetrahymena

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Regulation of Macronuclear DNA Content in *Tetrahymena thermophila**†

Riding The Ciliate Cell Cycle—A Thirty‐Five‐Year Prospective^a