Genomic Exclusion in Tetrahymena: Genetic Basis *

Genomic exclusion is an aberration that occurs during conjugation in variety 1 of Tetrahymena pyriformis. Instead of containing markers from both parents, the out cross pairs are either homozygous for all the genes of one parent (unilateral genomic exclusion); or, some of the pairs are homozygous for the genes of one parent and other pairs are homozygous for the genes of the other parent (bilateral genomic exclusion). This phenomenon was first demonstrated in the C strain: some stocks evoke unilateral genomic exclusion; others, bilateral genomic exclusion. C*, inbred for 5 generations, was used to explore this phenomenon in some detail since unilateral genomic exclusion of C genes occurs in almost all pairs in outcrosses of C*. In a mating of C*, both exconjugants are recovered, both are dipioid and similar in phenotype. Using morphological markers, C* can be shown to participate in the mating; therefore, C* does not induce illegitimate matings of the normal mate. When the normal mate is heterozygous for alleles ( H A / H D ) not present in C*, 3 classes of offspring ( H A / H A , H A / H D and H D / H D ) are produced in a 1:2:1 ratio. These observations indicate that 2 meiotic products of the normal mate unite to form the syn carya. The genetic ratios obtained in 1 and 2 factor crosses limit the possible cytogenetic bases for genomic exclusion. They suggest that 1 of the 4 haploid nuclei replicates and the replica fuses randomly with any 1 of the 4 nuclei. The 2 schemes of nuclear behavior (single fertilization, double fertilization) that would satisfy these requirements have not yet been resolved.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73138/1/j.1550-7408.1963.tb01699.x.pd

GENETIC CONTROL OF THE ESTERASES IN THE PROTOZOAN TETRAHYMENA PYRIFORMIS *

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72830/1/j.1749-6632.1961.tb35571.x.pd

Macronuclear persistence of sequences normally eliminated during development in Tetrahymena thermophila

During conjugation in the ciliated protozoan, Tetrahymena thermophila , a somatic MAC-ronucleus develops from the germinal MICronucleus. Ten to 20 percent of the MIC genome is eliminated during this process. Three repetitive families have been identified which have different levels of repetition in the MIC and are eliminated to different degrees in the MAC. Some members of two of these families persist in the MAC. In this study, we have looked at these persistent sequences in the MAC of cell lines from a variety of sources including several inbed strains, two sets of caryonides, caryonidal subclones, and vegetatively aged cell clones. The results suggest that the sequences that remain in the MAC have a genetic predisposition to persist. However, epigenetic variations occur as the MAC develops so that only some of the persistent sequences are actually observed in a particular MAC. Polymorphisms may be generated if alternative processing of a single MIC segment occurs. These polymorphisms can later be resolved by phenotypic assortment during vegetative growth. These facultatively persistent sequences appear to differ from sequences previously described in this organism.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50176/1/1020060205_ftp.pd

Effect of Acetate on Esterase C Activity During the Growth Cycle of Paramecium *

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/100130/1/j.1550-7408.1978.tb04415.x.pd

Alternative Processing of Sequences During Macronuclear Development in Tetrahymena thermophila 1

DNA is eliminated during development of the somatic MACronucleus from the germinal MICronucleus in the ciliated protozoan, Tetrahymena thermophila. Facultatively persistent sequences are a class of sequences that persist in the MAC DNA of some cell lines but are eliminated from the MAC DNA of other cell lines. One cloned MAC fragment contains a persistent sequence as well as sequences normally retained in the MAC. When this cloned fragment was used to construct MAC restriction maps of this region in cell lines whose MAC DNAs do, or do not, contain the persistent sequence, extensive variation in the map flanking this region was observed. The different DNA rearrangements of this MIC segment are epigenetically determined during or soon after MAC development. Moreover, different rearrangements may occur among the 45 copies of this MIC segment as a MAC is formed, resulting in polymorphisms that are later resolved by phenotypic assortment.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/74715/1/j.1550-7408.1986.tb05551.x.pd

Intraspecies Variability in the Esterases and Acid Phosphatases of Four Species of the Paramecium aurelia Complex 1

One hundred eighty-eight stocks of Paramecium primaurelia. P. biaurelia, P. tetraurelia. and P. octaurelia were grown axenically and screened for variation in four different esterases and acid phosphatase using starch gel electrophoresis. Major observations: frequency of intraspecies variation for these enzymes is much lower in these four species than in other organisms; hypervariability for two esterases occurs in P. biaurelia both in isolates from worldwide locales and in a restricted locale; clustering of variations occurs in a high proportion of variant stocks in all four species; frequency of intraspecies variation is highest in Central and South America for all four species; and geographical differentiation is lacking between stocks in the same species both for common as well as variant phenotypes despite the cosmopolitan distribution of these species. These results are not correlated with adaptations that favor inbreeding over outbreeding. nor is the possession of bacterial endosymbionts strongly correlated with enzyme variation. When the riequency of intraspecies variation was examined for the aurelia complex of species as a whole for 13 enzymes, mitochondrial DNA, and ribosomal DNA, differences between enzymes in frequency of variation could be seen, ranging from less than 2% for seven enzymes to 12.4% for glucosephosphate isomerase, a value similar to that observed for malic dehydrogenase, mitochondrial DNA, and ribosomal DNA in P. tetraurelia. The percentage of polymorphic enzyme loci in the complex as a whole was found to be much lower than that observed for other organisms. For the species more intensely studied in this paper the level of genetic polymorphism was also much lower, although P. biaurelia showed greater variability for two of the enzymes.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/71609/1/j.1550-7408.1983.tb01047.x.pd

Highly Purified Micro- and Macronuclei from Tetrahymena thermophila Isolated by Percoll Gradients 1

A new procedure is described that utilizes Percoll gradients for purifying micronuclei (MIC) and macronuclei (MAC) from Tetrahymena thermophila . Separation of MIC from MAC during certrifugation in Percoll gradients occurs as a result of their difference in size rather than density. Three kinds of tests were used to evaluate the purity of the nuclei: visualization of the nuclei by light microscopy; examination of the nuclei by electron microscopy; and Southern blots of MIC and MAC DNA probed with the 5s rRNA genes or a fragment from the MAC extrachromosomal rDNA molecule. When examined under the light microscope, the isolated MIC and MAC have much lower nuclear cross contamination levels than previous methods have reported. MIC's contaminated with less than 1 MAC in 1000 MIC and MAC's contaminated with less than 1 MIC in 500 MAC can be routinely prepared. Quantitative analyses of electron micrographs gave higher estimates of cross contamination in our purified nuclei, which may, in part, be explained by the difficulty in identifying small MIC or MAC fragments. Southern blots of MIC and MAC DNA probed with 5s rDNA confirmed the level of MAC contamination in the MIC estimated by light microscopy during purification of the nuclei. The level of nucleolar contamination in the MIC was estimated at 10% by Southern blots of MIC and MAC DNA, derived from a heterokaryon with distinctive MIC and MAC Bam HI sites, using an rDNA probe.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75629/1/j.1550-7408.1983.tb01027.x.pd

Esterase Variants in Four Species of the Paramecium aurelia Complex 1

One hundred eighty-eight stocks of Paramecium primaurelia, P. biaurelia, P. tetraurelia , and P. octaurelia were grown axenically and tested for five esterases, visualized by starch gel electrophoresis, in a search for variant stocks. The five esterases can be distinguished on the bases of their substrate specificity, sensitivity to an inhibitor, and response to different growth conditions. This paper addresses the nature of the electrophoretic change in mobility of the variant stocks in order that species relationships can be more accurately assessed. Crosses carried out in all four species show that single genes determine the differences in mobility between variant and common subtypes. Extracts of variant stocks that gave similar patterns were run against each other, tested for their sensitivity to the inhibitor, and the pattern was compared to that found in extracts of stocks with variant and common subtypes in other species. The majority of the variants in P. primaurelia, P. tetraurelia , and P. octaurelia show an electrophoretic mobility characteristic of a common subtype, or a variant, in another species. The same proportion of variant subtypes as common subtypes have mobilities similar to esterase subtypes found in other species. Of the four species examined in this paper, P. tetraurelia and P. octaurelia appear to be most closely related on the basis of shared esterase subtypes. In P. biaurelia the mobilities of most of the variants are unique, as are the common esterase subtypes in this species. P. biaurelia stands out as having the greatest number of esterase subtypes, with very few of them homologous to subtypes found in other species. This observation supports the idea of greater diversification of stocks within P. biaurelia than for the other three species.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75741/1/j.1550-7408.1982.tb01346.x.pd

Intersyngenic variations in the esterases of bacterized Paramecium aurelia

The esterase isozymes were surveyed in bacterized stocks representative of all 14 syngens of Paramecium aurelia by starch gel electrophoresis. The properties of substrate specificity and independent variation of particular isozymes permit the ordering of the differences observed among stocks. Differences can arise from several sources: bacterial variation, intrasyngenic variation, and intersyngenic variation. Bacterial esterases tend to be found in certain zonal areas (see Rowe et al. , 1971) and produce minor stock differences, which are erratic in their distribution. Unlike the situation found in Tetrahymena pyriformis , major intrasyngenic variations are rare in P. aurelia except in syngen 2. This lack of intrasyngenic variation is significant in view of the wide differences in geographic origin and micronuclear chromosome numbers among stocks within a syngen. It suggests that certain esterase genotypes must be under stringent selection within a syngen. The lack of intrasyngenic variation permits assessment of intersyngenic relationships. Syngens differ in a complex way from each other, suggesting that several gene differences may be involved. The syngens can be classified on the basis of their esterases. Syngens which have been shown to be more closely related in terms of cross-mating, breeding systems, and other criteria tend to be more similar in their esterase isozymes. The isozyme technique confirms relationships previously suggested among syngens and offers the promise of eventual assessment of evolutionary distances among syngens. However, establishment of these relationships will be clearer in the absence of bacteria.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/44169/1/10528_2004_Article_BF00485641.pd

Eliminated sequences with different copy numbers clustered in the micronuclear genome of Tetrahymena thermophila

As the ciliated protozoan Tetrahymena thermophila develops a new macronucleus (MAC) from products of its micronucleus (MIC), several repetitive sequences are eliminated from the MAC genome. Four MIC DNA clones containing repetitive sequences that are eliminated from the MAC were obtained. One clone contains a representative from each of three families of eliminated sequences. One, present in 200–300 copies in the MIC, is almost completely eliminated from the MAC. A second, present in approximately 50 copies in the MIC, is scattered throughout the genome, although up to half of the family members examined could be localized to chromosome 2. Approximately one tenth of the members of this less repetitive family persist in the MAC while the rest are eliminated. The third type of eliminated sequence has three to four members, all of which are eliminated from the MAC. Three of the members are located on three of the five MIC chromosomes, and one could not be mapped. This sequence is clustered with the other two families of sequences in at least three of the four sites. All three types of eliminated sequences are found in similar arrangements in the MIC of several different inbred strains of T. thermophila .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47561/1/438_2004_Article_BF00397988.pd