scholarly journals Heat synchronization of cell division in Tetrahymena thermophila and a mutant, NP1, with a temperature sensitive defect for oral development

1978 ◽  
Vol 43 (4) ◽  
pp. 255-263 ◽  
Author(s):  
Peter B. Suhr-Jessen
Keyword(s):  
Cell Division ◽  
Tetrahymena Thermophila ◽  
Temperature Sensitive ◽  
Oral Development ◽  
Synchronization Of Cell Division

scholarly journals The relationship between the excess-delay phenomenon and temperature-sensitive periods in Tetrahymena thermophila

1980 ◽  
Vol 43 (1) ◽  
pp. 75-91
Author(s):  
J. Frankel ◽  
J. Mohler ◽  
A.K. Frankel
Keyword(s):  
Cell Division ◽  
Transition Point ◽  
Tetrahymena Thermophila ◽  
Characteristic Parameter ◽  
Sensitive Period ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Heat Shocks ◽  
Delay Phenomenon ◽  
The Relationship

Although temperatures of 37.5 and 39 degrees C allow continuous and rapid exponential growth of wild type Tetrahymena thermophila, sudden shifts up to these temperatures can bring about long excess-delays of cell division with accompanying resorption of developing oral primordia. A characteristic parameter of this delay-phenomenon is the physiological transition point, before which delays are maximal and after which they are negligible. When measured at a restrictive temperature that does not induce excess delays (36 degrees C), the end of the temperature-sensitive period of the cell division arrest of mutant cdaA1 precedes the physiological transition point, that of cdaH1 roughly coincides with it, while the entire temperature-sensitive period of cdaC2 comes after the physiological transition point. When cdaA1 cells are exposed to 37.5 degrees C or above, the manifestations of temperature sensitivity are drastically affected: the estimate of the end of the temperature-sensitive period (the execution point) becomes spuriously late, and the characteristic division arrest following heat shocks is not manifested. The differential effects of the higher restrictive temperatures on cdaH1 are most subtle, whereas those on cdaC2 are negligible. We conclude that the excess-delay phenomenon involves a set-back of genemediated processes occurring at specific stages of the cell cycle.

Temperature-sensitive periods of mutations affecting cell division in Tetrahymena thermophila

1980 ◽  
Vol 43 (1) ◽  
pp. 59-74 ◽  
Author(s):  
J. Frankel ◽  
J. Mohler ◽  
A.K. Frankel
Keyword(s):  
Cell Division ◽  
Tetrahymena Thermophila ◽  
Sensitive Period ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Permissive Temperature ◽  
Physiological Effects ◽  
Sensitive Periods ◽  
Phenotypic Abnormality ◽  
Mutant Cells

Temperature-sensitive periods were determined by application of temperature shifts and shocks to 3 temperature-sensitive cell division arrest (cda) mutants of Tetrahymena thermophila. A restrictive temperature, 36 degrees C, was found at which all 3 mutants are fully penetrant, yet other physiological effects are minimal. At this temperature, the temperature-sensitive period of cdaC2 is a unique 5-min period in mid-division, that of cdaA1 is a similarly brief period situated about 0.5 h prior to cell division, while the temperature-sensitive period of cdaH1 is 20 to 30 min long and immediately precedes cell division. These periods either coincide with (cdaC2, cdaH1) or immediately precede (cdaA1) the onset of phenotypic abnormality at the restrictive temperature. Brief exposure to 36 degrees C during the temperature-sensitive period in any of these mutants brings about irreversible arrest of division furrows in progress or preparation. Mutant cells suffering such arrest can, however, divide again at a permissive temperature by forming new furrows at different sites.

scholarly journals MUTANTS OF TETRAHYMENA THERMOPHILA WITH TEMPERATURE-SENSITIVE FOOD VACUOLE FORMATION. I. ISOLATION AND GENETIC CHARACTERIZATION

Genetics ◽  
1979 ◽  
Vol 92 (4) ◽  
pp. 1061-1077
Author(s):  
Peter B Suhr-Jessen ◽  
Eduardo Orias
Keyword(s):  
Tetrahymena Thermophila ◽  
Germ Line ◽  
Complementation Group ◽  
Food Vacuole ◽  
Temperature Sensitive ◽  
Genetic Dissection ◽  
Food Vacuoles ◽  
Per Se ◽  
Oral Development

ABSTRACT Germ-line mutants have been isolated in Tetrahymena thermophila that have recessive, temperature-sensitive defects in phagocytosis. Nitrosoguanidine-mutagenized cells were induced to undergo cytogamy, and clones were isolated that were unable to form food vacuoles after two days of growth at 39°. Most of the mutants belong to a single complementation group, designated vacA. They have defects in oral development—not in phagocytosis per se—that are undetectable under light microscopy. One fertile mutant, phenotypically indistinguishable from the vacA group, has its vac mutation(s) restricted to the macronucleus, and it is a heterokaryon for two other markers. This clone probably resulted from a failure of the two gametic nuclei to fuse after nor,mal exchange. Two additional mutants were studied, but their sterility prevented a full genetic analysis. One of these clones has a rudimentary oral apparatus and defective contractile vacuole pores; both defects may be determined by the same mutation. The other clone has a structurally normal oral apparatus and may be defective in phagocytosis per se.——The induction and characterization of germ-line mutations that affect oral development open the way for the genetic dissection of the morphogenesis of a complex eukaryotic organelle, and make available additional useful mutants for the study of nutrition and transmembrane active transport.

scholarly journals Causal relations among cell cycle processes in Tetrahymena pyriformis. An analysis employing temperature-sensitive mutants.

1976 ◽  
Vol 71 (1) ◽  
pp. 242-260 ◽  
Author(s):  
J Frankel ◽  
L M Jenkins ◽  
L E DeBault
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Dna Synthesis ◽  
Tetrahymena Pyriformis ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Developmental Pathway ◽  
Temperature Sensitive Mutants ◽  
Oral Development ◽  
Macronuclear Division

Utilization of temperature-sensitive mutants of Tetrahymena pyriformis affected in cell division or developmental pathway selection has permitted elucidation of causal dependencies interrelating micronuclear and macronuclear replication and division, oral development, and cytokinesis. In those mutants in which cell division is specifically blocked at restrictive temperatures, micronuclear division proceeds with somewhat accelerated periodicity but maintains normal coupling to predivision oral development. Macronuclear division is almost totally suppressed in an early acting mutant (mola) that prevents formation of the fission zone, and is variably affected in other mutants (such as mo3) that allow the fission zone to form but arrest constriction. However, macronuclear DNA synthesis can proceed for about four cycles in the nondividing mutant cells. A second class of mutants (psm) undergoes a switch of developmental pathway such that cells fail to enter division but instead repeatedly carry out an unusual type of oral replacement while growing in nutrient medium at the restrictive temperature. Under these circumstances no nuclei divide, yet macronuclear DNA accumulation continues. These results suggest that (a) macronuclear division is stringently affected by restriction of cell division, (b) micronuclear division and replication can continue in cells that are undergoing the type of oral development that is characteristic of division cycles, and (c) macronuclear DNA synthesis can continue in growing cells regardless of their developmental status. The observed relationships among events are consistent with the further suggestion that the cell cycle in this organism may consist of separate clusters of events. with a varying degree of coupling among clusters. A minimal model of the Tetrahymena cell cycle that takes these phenomena into account is suggested.

scholarly journals THE SELECTION OF S. CEREVISIAE MUTANTS DEFECTIVE IN THE START EVENT OF CELL DIVISION

Genetics ◽  
1980 ◽  
Vol 95 (3) ◽  
pp. 561-577 ◽  
Author(s):  
Steven I Reed
Keyword(s):  
Cell Division ◽  
Genetic Analysis ◽  
Linkage Map ◽  
Nuclear Genes ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Mating Pheromone ◽  
Preliminary Characterization ◽  
Complementation Groups ◽  
Selection Of

ABSTRACT Thirty-three temperature-sensitive mutations defective in the start event of the cell division cycle of Saccharomyces cereuisiae were isolated and subjected to preliminary characterization. Complementation studies assigned these mutations to four complementation groups, one of which, cdc28, has been described previously. Genetic analysis revealed that these complementation groups define single nuclear genes, unlinked to one another. One of the three newly identified genes, cdc37, has been located in the yeast linkage map on chromosome IV, two meiotic map units distal to hom2.—Each mutation produces stage-specific arrest of cell division at start, the same point where mating pheromone interrupts division. After synchronization at start by incubation at the restrictive temperature, the mutants retain the capacity to enlarge and to conjugate.

Directionality of microtubule assembly: an in vivo study with the ciliate Tetrahymena

1978 ◽  
Vol 33 (1) ◽  
pp. 227-234
Author(s):  
S.F. Ng
Keyword(s):  
Tetrahymena Thermophila ◽  
Recessive Gene ◽  
Microtubule Assembly ◽  
In Vivo Study ◽  
Temperature Sensitive ◽  
Sensitive Mutant ◽  
Posterior Portion ◽  
Temperature Sensitive Mutant

A temperature-sensitive mutant homozygous for the recessive gene molb in Tetrahymena thermophila offers opportunity for studying the direction of microtubule assembly in vivo. At 39 degrees C the mutant fails to divide properly; the 2 daughter animals remain attached and bend over each other. As revealed by protargol staining, the bending results in acute turning and breaking of some of the longitudinal microtubular bands close and parallel to the surface. Hence, 2 broken microtubular ends are available for study of the problem of directionality of microtubule assembly, by assessing which of the 2 ends regenerates. In most cases the posterior portion of the longitudinal microtubular band regenerates. The present study hence supports the conclusion based on in vitro observation in other systems that microtubule assembly is predominantly unidirectional.

Test for temporal or spatial restrictions in gene product function during the cell division cycle

1983 ◽  
Vol 3 (7) ◽  
pp. 1255-1265
Author(s):  
S K Dutcher ◽  
L H Hartwell
Keyword(s):  
Cell Division ◽  
Gene Product ◽  
Gene Mutations ◽  
Cell Division Cycle ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Product Function ◽  
Complete Cell ◽  
Relationship Of ◽  
Gene Product Function

The ability of a functional gene to complement a nonfunctional gene may depend upon the intracellular relationship of the two genes. If so, the function of the gene product in question must be limited in time or in space. CDC (cell division cycle) gene products of Saccharomyces cerevisiae control discrete steps in cell division; therefore, they constitute reasonable candidates for genes that function with temporal or spatial restrictions. In an attempt to reveal such restrictions, we compared the ability of a CDC gene to complement a temperature-sensitive cdc gene in diploids where the genes are located within the same nucleus to complementation in heterokaryons where the genes are located in different nuclei. In CDC X cdc matings, complementation was monitored in rare heterokaryons by assaying the production of cdc haploid progeny (cytoductants) at the restrictive temperature. The production of cdc cytoductants indicates that the cdc nucleus was able to complete cell division at the restrictive temperature and implies that the CDC gene product was provided by the other nucleus or by cytoplasm in the heterokaryon. Cytoductants from cdc28 or cdc37 crosses were not efficiently produced, suggesting that these two genes are restricted spatially or temporally in their function. We found that of the cdc mutants tested 33 were complemented; cdc cytoductants were recovered at least as frequently as CDC cytoductants. A particularly interesting example was provided by the CDC4 gene. Mutations in CDC4 were found previously to produce a defect in both cell division and karyogamy. Surprisingly, the cell division defect of cdc4 nuclei is complemented by CDC4 nuclei in a heterokaryon, whereas the karyogamy defect is not.

scholarly journals Cell cycle arrest of cdc mutants and specificity of the RAD9 checkpoint.

Genetics ◽  
1993 ◽  
Vol 134 (1) ◽  
pp. 63-80 ◽  
Author(s):  
T A Weinert ◽  
L H Hartwell
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Dna Replication ◽  
Cell Cycle Control ◽  
Dna Lesions ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
A Cell ◽  
G2 Phase ◽  
Rad Mutants

Abstract In eucaryotes a cell cycle control called a checkpoint ensures that mitosis occurs only after chromosomes are completely replicated and any damage is repaired. The function of this checkpoint in budding yeast requires the RAD9 gene. Here we examine the role of the RAD9 gene in the arrest of the 12 cell division cycle (cdc) mutants, temperature-sensitive lethal mutants that arrest in specific phases of the cell cycle at a restrictive temperature. We found that in four cdc mutants the cdc rad9 cells failed to arrest after a shift to the restrictive temperature, rather they continued cell division and died rapidly, whereas the cdc RAD cells arrested and remained viable. The cell cycle and genetic phenotypes of the 12 cdc RAD mutants indicate the function of the RAD9 checkpoint is phase-specific and signal-specific. First, the four cdc RAD mutants that required RAD9 each arrested in the late S/G2 phase after a shift to the restrictive temperature when DNA replication was complete or nearly complete, and second, each leaves DNA lesions when the CDC gene product is limiting for cell division. Three of the four CDC genes are known to encode DNA replication enzymes. We found that the RAD17 gene is also essential for the function of the RAD9 checkpoint because it is required for phase-specific arrest of the same four cdc mutants. We also show that both X- or UV-irradiated cells require the RAD9 and RAD17 genes for delay in the G2 phase. Together, these results indicate that the RAD9 checkpoint is apparently activated only by DNA lesions and arrests cell division only in the late S/G2 phase.

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