scholarly journals Analysis of flagellar size control using a mutant of Chlamydomonas reinhardtii with a variable number of flagella.

1982 ◽  
Vol 92 (1) ◽  
pp. 170-175 ◽  
Author(s):  
M R Kuchka ◽  
J W Jarvik
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Cell Size ◽  
Size Control ◽  
Variable Number ◽  
Pool Size ◽  
Precursor Protein ◽  
Wild Type ◽  
Protein Pool ◽  
Mutant Cells ◽  
Pool Sizes

A mutant of Chlamydomonas reinhardtii with a variable number of flagella per cell has been used to investigate flagellar size control. The mutant and wild-type do not differ in cell size nor in flagellar length, yet the size of the intracellular pool of flagellar precursor protein can differ dramatically among individual mutant cells, with, for example, triflagellate cells having three times the pool of monoflagellate cells. Because cells of the same size, but with very different pool sizes, have flagella of identical length, it appears that the concentration of the unassembled flagellar precursor protein pool does not regulate flagellar length. The relation between cell size, pool size, and flagellar length has also been investigated for wild-type cells of different sizes and ploidies. Again, flagellar length appears to be maintained independent of pool size or concentration.

The kinetics of the B cyclin p56cdc13 and the phosphatase p80cdc25 during the cell cycle of the fission yeast Schizosaccharomyces pombe

1996 ◽  
Vol 109 (6) ◽  
pp. 1647-1653 ◽  
Author(s):  
J. Creanor ◽  
J.M. Mitchison
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Mammalian Cells ◽  
Size Control ◽  
Base Line ◽  
Wild Type ◽  
Main Band ◽  
The Times ◽  
G1 Cells ◽  
Mutant Cells ◽  
Kinetics Of

The levels of the B cyclin p56cdc13 and the phosphatase p80cdc25 have been followed in selection-synchronised cultures of Schizosaccharomyces pombe wild-type and wee1 mutant cells. p56cdc13 has also been followed in induction-synchronised cells of the mutant cdc2-33. The main conclusions are: (1) cdc13 levels in wild-type cells start to rise from base line at about mid-G2, reach a peak before mitosis and then fall slowly through G1. Cells exit mitosis with appreciable levels of cdc13. (2) cdc13 levels in wee1 cells fall to zero in interphase. They also start to rise at the beginning of G2, which may be related to the absence of a mitotic size control. (3) cdc25 starts to rise later and reaches a peak after mitosis. This is not what would be expected from a simple mitotic inducer and suggests that cdc25 has an important function at the end of mitosis. (4) An upper (heavier) band of cdc25 peaks at the same time as the main band but rises and falls more rapidly. If this is a hyperphosphorylated form, its timing shows that it is most unlikely to function in the ways shown for such a form in eggs and mammalian cells. (5) Experiments with the mutant cdc10-129 and with hydroxyurea show that the initial signal to begin synthesis of cdc13 originates at Start. (6) In induction synchrony, where G2 spans across cell division, there is evidence that some events in one cycle cannot start in the previous one. (7) Revised timings are given for the times of mitosis in these cultures.

Growth in cell length in the fission yeast Schizosaccharomyces pombe

1985 ◽  
Vol 75 (1) ◽  
pp. 357-376 ◽  
Author(s):  
J.M. Mitchison ◽  
P. Nurse
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Cell Size ◽  
Single Cells ◽  
Temperature Shift ◽  
Time Lapse ◽  
Cell Length ◽  
Wild Type ◽  
Cycle Growth ◽  
A Cell ◽  
Mutant Cells

The cylindrical cells of Schizosaccharomyces pombe grow in length by extension at the ends and not the middle. At the beginning of the cell cycle, growth is restricted to the ‘old end’, which existed in the previous cycle. Later on, the ‘new end’, formed from the septum, starts to grow at a point in the cycle that we have called NETO (‘new end take-off’). Fluorescence microscopy on cells stained with Calcofluor has been used to study NETO in size mutants, in blocked cdc mutants and with different growth temperatures and media. In wild-type cells (strain 972) NETO happens at 0.34 of the cycle with a cell length of 9.5 microns. With size mutants that are smaller at division, NETO takes place at the same size (9.0-9.5 microns) but this is not achieved until later in the cycle. Another control operates in larger size mutants since NETO occurs at the same stage of the cycle (about 0.32) as in wild type but at a larger cell size. This control is probably a requirement to have completed an event in early G2, since most cdc mutant cells blocked before this point in the cycle do not show NETO whereas most of those blocked in late G2 do show it. We conclude that NETO only happens if: (1) the cell length is greater than a critical value of 9.0-9.5 microns; and (2) the cell has traversed the first 0.3-0.35 of the cycle and passed early G2. NETO is delayed in poor media, in which cell size is also reduced. Temperature has little effect on NETO under steady-state conditions, but there is a transient delay for some hours after a temperature shift. NETO is later in another wild-type strain, 132. Time-lapse photomicrography was used to follow the rates of length growth in single cells. Wild-type cells showed two linear segments during the first 75% of the cycle. There was a rate-change point (RCP), coincident with NETO, where the rate of total length extension increased by 35%. This increase was not due simply to the start of new-end growth, since old-end growth slowed down in some cells at the RCP. cdc 11.123 is a mutant in which septation and division is blocked at 35 degrees C but nuclear division continues.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Cell-size regulation in budding yeast does not depend on linear accumulation of Whi5

2020 ◽  
Vol 117 (25) ◽  
pp. 14243-14250 ◽  
Author(s):  
Felix Barber ◽  
Ariel Amir ◽  
Andrew W. Murray
Keyword(s):  
Cell Cycle ◽  
Cell Size ◽  
Cell Cycle Regulation ◽  
Budding Yeast ◽  
Size Control ◽  
Size Distributions ◽  
Wild Type ◽  
Gal1 Promoter ◽  
The Mean ◽  
Cycle Regulation

Cells must couple cell-cycle progress to their growth rate to restrict the spread of cell sizes present throughout a population. Linear, rather than exponential, accumulation of Whi5, was proposed to provide this coordination by causing a higher Whi5 concentration in cells born at a smaller size. We tested this model using the inducibleGAL1promoter to make the Whi5 concentration independent of cell size. At an expression level that equalizes the mean cell size with that of wild-type cells, the size distributions of cells with galactose-induced Whi5 expression and wild-type cells are indistinguishable. Fluorescence microscopy confirms that the endogenous andGAL1promoters produce different relationships between Whi5 concentration and cell volume without diminishing size control in the G1 phase. We also expressed Cln3 from the GAL1 promoter, finding that the spread in cell sizes for an asynchronous population is unaffected by this perturbation. Our findings indicate that size control in budding yeast does not fundamentally originate from the linear accumulation of Whi5, contradicting a previous claim and demonstrating the need for further models of cell-cycle regulation to explain how cell size controls passage through Start.

scholarly journals Thecdr2+Gene Encodes a Regulator of G2/M Progression and Cytokinesis inSchizosaccharomyces pombe

1998 ◽  
Vol 9 (12) ◽  
pp. 3399-3415 ◽  
Author(s):  
Connie S. Breeding ◽  
James Hudson ◽  
Mohan K. Balasubramanian ◽  
Sean M. Hemmingsen ◽  
Paul G. Young ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Cell Size ◽  
Size Control ◽  
S Phase ◽  
Nutrient Deprivation ◽  
Cyclin Dependent Kinase ◽  
Wild Type ◽  
M Cell ◽  
Mitotic Control

Schizosaccharomyces pombe cells respond to nutrient deprivation by altering G2/M cell size control. The G2/M transition is controlled by activation of the cyclin-dependent kinase Cdc2p. Cdc2p activation is regulated both positively and negatively. cdr2+was identified in a screen for regulators of mitotic control during nutrient deprivation. We have cloned cdr2+and have found that it encodes a putative serine-threonine protein kinase that is related to Saccharomyces cerevisiae Gin4p and S. pombe Cdr1p/Nim1p.cdr2+is not essential for viability, but cells lacking cdr2+are elongated relative to wild-type cells, spending a longer period of time in G2. Because of this property, upon nitrogen deprivationcdr2+mutants do not arrest in G1, but rather undergo another round of S phase and arrest in G2from which they are able to enter a state of quiescence. Genetic evidence suggests thatcdr2+acts as a mitotic inducer, functioning through wee1+, and is also important for the completion of cytokinesis at 36°C. Defects in cytokinesis are also generated by the overproduction of Cdr2p, but these defects are independent of wee1+, suggesting thatcdr2+encodes a second activity involved in cytokinesis.

scholarly journals Cell Structure Changes in the Hyperthermophilic Crenarchaeon Sulfolobus islandicus Lacking the S-Layer

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Changyi Zhang ◽  
Rebecca L. Wipfler ◽  
Yuan Li ◽  
Zhiyu Wang ◽  
Emily N. Hallett ◽  
...  
Keyword(s):  
Cell Division ◽  
Cell Size ◽  
Cell Fusion ◽  
Cell Biology ◽  
Cell Structure ◽  
Evolutionary Relationship ◽  
Wild Type ◽  
Sulfolobus Islandicus ◽  
Mutant Cells

ABSTRACT Rediscovery of the ancient evolutionary relationship between archaea and eukaryotes has revitalized interest in archaeal cell biology. Key to the understanding of archaeal cells is the surface layer (S-layer), which is commonly found in Archaea but whose in vivo function is unknown. Here, we investigate the architecture and cellular roles of the S-layer in the hyperthermophilic crenarchaeon Sulfolobus islandicus. Electron micrographs of mutant cells lacking slaA or both slaA and slaB confirm the absence of the outermost layer (SlaA), whereas cells with intact or partially or completely detached SlaA are observed for the ΔslaB mutant. We experimentally identify a novel S-layer-associated protein, M164_1049, which does not functionally replace its homolog SlaB but likely assists SlaB to stabilize SlaA. Mutants deficient in the SlaA outer layer form large cell aggregates, and individual cell size varies, increasing significantly up to six times the diameter of wild-type cells. We show that the ΔslaA mutant cells exhibit more sensitivity to hyperosmotic stress but are not reduced to wild-type cell size. The ΔslaA mutant contains aberrant chromosome copy numbers not seen in wild-type cells, in which the cell cycle is tightly regulated. Together, these data suggest that the lack of SlaA results in either cell fusion or irregularities in cell division. Our studies show the key physiological and cellular functions of the S-layer in this archaeal cell. IMPORTANCE The S-layer is considered to be the sole component of the cell wall in Sulfolobales, a taxonomic group within the Crenarchaeota whose cellular features have been suggested to have a close relationship to the last archaea-eukaryote common ancestor. In this study, we genetically dissect how the two previously characterized S-layer genes as well as a newly identified S-layer-associated protein-encoding gene contribute to the S-layer architecture in Sulfolobus. We provide genetic evidence for the first time showing that the slaA gene is a key cell morphology determinant and may play a role in Sulfolobus cell division or/and cell fusion.

Ultrastructure of Chlamydomonas reinhardtii following exposure to paraquat: comparison of wild type and a paraquat-resistant mutant

1993 ◽  
Vol 71 (1) ◽  
pp. 174-182 ◽  
Author(s):  
Douglas F. Bray ◽  
John R. Bagu ◽  
Kazuo Nakamura
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Wild Type Strain ◽  
Single Gene ◽  
Resistant Mutant ◽  
Cross Resistance ◽  
Wild Type ◽  
Ultrastructural Examination ◽  
Unicellular Green Alga ◽  
Strain 137C ◽  
Mutant Cells

A mutant (NL-51) of the unicellular green alga Chlamydomonas reinhardtii Dangeard isolated from a wild-type strain (137c+) was shown to be resistant to the bipyridilium herbicide paraquat at the concentration at which growth of the wild type was inhibited. Tetrad analysis from a cross between the mutant and the wild type showed 2:2 segregation, indicating that the resistance is under control of a single gene. Cross-resistance of the mutant to methionine and to methionine combined with riboflavin suggested that the resistance is due to increased levels of one of the enzymes capable of detoxifying active oxygens. Ultrastructural examination of mutant and wild-type cells exposed to paraquat revealed that the mutant cells were 3 to 4 times more resistant, but both strains showed the same sequence of deterioration. Damage was first manifested as swelling of the mitochondria and dilation of the perinuclear space. This was followed by disintegration of the nuclear matrix and the chloroplast thylakoids. Key words: Chlamydomonas reinhardtii, methionine resistance, paraquat, paraquat-resistant mutant, ultrastructure.

scholarly journals Asymmetric properties of the Chlamydomonas reinhardtii cytoskeleton direct rhodopsin photoreceptor localization

2011 ◽  
Vol 193 (4) ◽  
pp. 741-753 ◽  
Author(s):  
Telsa M. Mittelmeier ◽  
Joseph S. Boyd ◽  
Mary Rose Lamb ◽  
Carol L. Dieckmann
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Basal Body ◽  
Cytoskeletal Protein ◽  
Wild Type ◽  
Unicellular Green Alga ◽  
Protein Mutations ◽  
The Relationship ◽  
Mutant Cells ◽  
In Cells

The eyespot of the unicellular green alga Chlamydomonas reinhardtii is a photoreceptive organelle required for phototaxis. Relative to the anterior flagella, the eyespot is asymmetrically positioned adjacent to the daughter four-membered rootlet (D4), a unique bundle of acetylated microtubules extending from the daughter basal body toward the posterior of the cell. Here, we detail the relationship between the rhodopsin eyespot photoreceptor Channelrhodopsin 1 (ChR1) and acetylated microtubules. In wild-type cells, ChR1 was observed in an equatorial patch adjacent to D4 near the end of the acetylated microtubules and along the D4 rootlet. In cells with cytoskeletal protein mutations, supernumerary ChR1 patches remained adjacent to acetylated microtubules. In mlt1 (multieyed) mutant cells, supernumerary photoreceptor patches were not restricted to the D4 rootlet, and more anterior eyespots correlated with shorter acetylated microtubule rootlets. The data suggest a model in which photoreceptor localization is dependent on microtubule-based trafficking selective for the D4 rootlet, which is perturbed in mlt1 mutant cells.

scholarly journals Triacylglyceride Production and Autophagous Responses in Chlamydomonas reinhardtii Depend on Resource Allocation and Carbon Source

2014 ◽  
Vol 13 (3) ◽  
pp. 392-400 ◽  
Author(s):  
Matthew P. Davey ◽  
Irmtraud Horst ◽  
Giang-Huong Duong ◽  
Eleanor V. Tomsett ◽  
Alexander C. P. Litvinenko ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Membrane Lipids ◽  
N Availability ◽  
Wild Type ◽  
Metabolic Switch ◽  
Content Type ◽  
Carbon Supply ◽  
Microalgal Biodiesel ◽  
In The Wild ◽  
Mutant Cells

ABSTRACT To improve the economic viability of microalgal biodiesel, it will be essential to optimize the productivity of fuel molecules such as triacylglyceride (TAG) within the microalgal cell. To understand some of the triggers required for the metabolic switch to TAG production, we studied the effect of the carbon supply (acetate or CO 2 ) in Chlamydomonas reinhardtii (wild type and the starchless sta6 mutant) grown under low N availability. As expected, initial rates of TAG production were much higher when acetate was present than under strictly photosynthetic conditions, particularly for the sta6 mutant, which cannot allocate resources to starch. However, in both strains, TAG production plateaued after a few days in mixotrophic cultures, whereas under autotrophic conditions, TAG levels continued to rise. Moreover, the reduced growth of the sta6 mutant meant that the greatest productivity (measured as mg TAG liter −1 day −1 ) was found in the wild type growing autotrophically. Wild-type cells responded to low N by autophagy, as shown by degradation of polar (membrane) lipids and loss of photosynthetic pigments, and this was less in cells supplied with acetate. In contrast, little or no autophagy was observed in sta6 mutant cells, regardless of the carbon supply. Instead, very high levels of free fatty acids were observed in the sta6 mutant, suggesting considerable alteration in metabolism. These measurements show the importance of carbon supply and strain selection for lipid productivity. Our findings will be of use for industrial cultivation, where it will be preferable to use fast-growing wild-type strains supplied with gaseous CO 2 under autotrophic conditions rather than require an exogenous supply of organic carbon.

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