Molecular evolution in hypotrichous ciliates: Sequence of the small subunit ribosomal RNA genes fromOnychodromus quadricornutus andOxytricha granulifera (Oxytrichidae,Hypotrichida, Ciliophora)

Summary The small subunit ribosomal RNA (16S-like rRNA) coding regions of the hypotrichous ciliatesOnychodromus quadricornutus andOxytricha granulifera were amplified using polymerase chain reaction techniques. Complete sequences were determined for the amplified genes and compared to those of other ciliated protozoa. In phylogenetic trees inferred using distance matrix methods oxytrichids are not seen as a cohesive phylogenetic group.Oxytricha nova is most closely related toStylonychia pustulata in a lineage that also includesO. quadricornutus. This phylogeny contradicts phylogenetic schemes in whichOnychodromus is considered to be a primitive hypotrichous ciliate and suggests thatO. nova was misidentified as members of the genusOxytricha.     

Thermodynamic and electrostatic properties of ternary Oxytricha nova TEBP-DNA complex

Telomeres constitute the nucleoprotein ends of eukaryotic chromosomes which are essential for their proper function. Telomere end binding protein (TEBP) from Oxytricha nova was among the first telomeric proteins, which were well characterized biologically. TEBP consists of two protein subunits (alpha, beta) and forms a ternary complex with single stranded telomeric DNA containing tandem repeats TTTTGGGG. This work presents the characterization of the thermodynamic and electrostatic properties of this complex by computational chemistry methods (continuum Poisson-Boltzmann and solvent accessible surface calculations). Our calculations give a new insight into molecular properties of studied system. Based on the thermodynamic analysis we provide a rationale for the experimental observation that alpha and ssDNA forms a binary complex and the beta subunit joins alpha:ssDNA complex only after the latter is formed. Calculations of distribution of the molecular electrostatic potential for protein subunits alone and for all possible binary complexes revealed the important role of the "guiding funnel" potential generated by alpha:ssDNA complex. This potential may help the beta subunit to dock to the already formed alpha:DNA intermediate in highly steric and electrostatic favorable manner. Our pK(a) calculations of TEBP are able to explain the experimental mobility shifts of the complex in electrophoretic non-denaturating gels.     

Commitment to Division in Ciliate Cell Cycles

Near the end of the cell cycle, ciliates commit irreversibly to cell division. The point of commitment occurs at the time of oral polykinetid assembly and micronuclear anaphase. The commitment is a checkpoint which requisites a threshold cell mass/ DNA ratio and stomatogenesis. It is also a physiological transition point, involving cdk protein kinases similar to those of other eukaryotes. Both P34 kD and P36 kD kinases, similar to the S. pombe cdc2 kinases, have been described to have activity as monomers. Subsequent to commitment to division, dramatic cytoskeletal modifications occur for separation of organelles, cortex morphogenesis and cytokinesis. Numerous mutants affecting cytoskeletal function associated with the division process have been obtained in several species. Of these, only the ccl mutant in Paramecium affects cell cycle progression prior to commitment to division. The material reviewed is used to speculate about the mechanisms of regulation of pre-fission morphogenesis and cell division related processes in ciliates.     

Relationships between Cell Cycle and Conjugation in 3 Hypotrichs*

SYNOPSIS. Relationships between the cell cycle and the beginning of conjugation were analyzed for 3 hypotrichs: Diophrys scutum, Oxytricha bifaria, and Euplotes crassus. The first 2 species enter conjugation with micronuclei in G₁; the latter species with a micronucleus in G₂. The 1st micronuclear division of conjugating E. crassus is mitotic. Thus meiotic DNA replication occurs when the cells of each species have already entered the mating process. Cells from asynchronous populations start conjugation with their macronuclei primarily in G₁ or more rarely at the beginning of the S stage in a percentage significantly different from that expected on the basis of random mating among all cells in the population. Also, macronuclear replication, when already begun, was blocked in cells undergoing conjugation. Therefore only the G₁ or the very early S stages of the cell cycle are compatible with conjugation in the 3 analyzed species.     

The Structure and Development of the Dorsal Bristle Complex of Oxytricha fallax and Stylonychia pustulata*

SYNOPSIS. Oxytricha fallax and Stylonychia pustulata possess 6 rows of dorsal bristle units. Each dorsal bristle unit consists of a pair of kinetosomes; the anterior kinetosome has a cilium and the posterior kinetosome a ciliary stub. The kinetosome pair, located at the bottom of a cortical pit surrounding the cilium and ciliary stub, is surrounded by an asymmetrical fibrillar mass. Future rows 1-4 are formed from 2 sets of primordia originating within mature dorsal rows 1-3. Rows 5 and 6 originate from the anterior regions of both right marginal cirral primordia. Old dorsal bristle units utilized in formation of primordia are presumably maintained in the new rows of the proter and opisthe; those outside the primordia are resorbed. The morphogenetic pattern of the Oxytrichidae is similar to those of the Urostylidae and Holostichidae, but quite different from that of the Euplotidae.     

阔口尖毛虫皮层纤毛器微管在不同生理条件下的分化

应用激光扫描共聚焦显微术,显示腹毛类纤毛虫阔口尖毛虫(Oxytricha platystoma)无性生殖过程中,新的口围带、波动膜、额腹横棘毛、左右缘棘毛微管先后分化,老纤毛器微管去分化,细胞分裂产生各含一套纤毛器微管的前、后两仔虫;生理改组过程中,口围带、波动膜、额腹横棘毛、左右缘棘毛微管发生去分化和再分化,细胞皮层微管胞器更新形成含一套纤毛器微管的新细胞。结果表明阔口尖毛虫在无性生殖和生理改组这两种不同的生理条件下,其纤毛器微管结构的形成或更新可能具有相同的细胞调控机制,形态发生中老纤毛器结构可能对新结构的发生和发育具有诱导定位和物质贡献的作用。     

Characterization and taxonomic validity of the ciliate Oxytricha trifallax (Class Spirotrichea) based on multiple gene sequences: limitations in identifying genera solely by morphology

Oxytricha trifallax - an established model organism for studying genome rearrangements, chromosome structure, scrambled genes, RNA-mediated epigenetic inheritance, and other phenomena - has been the subject of a nomenclature controversy for several years. Originally isolated as a sibling species of O. fallax, O. trifallax was reclassified in 1999 as Sterkiella histriomuscorum, a previously identified species, based on morphological similarity. The proper identification of O. trifallax is crucial to resolve in order to prevent confusion in both the comparative genomics and the general scientific communities. We analyzed nine conserved nuclear gene sequences between the two given species and several related ciliates. Phylogenetic analyses suggest that O. trifallax and a bona fide S. histriomuscorum have accumulated significant evolutionary divergence from each other relative to other ciliates such that they should be unequivocally classified as separate species. We also describe the original isolation of O. trifallax, including its comparison to O. fallax, and we provide criteria to identify future isolates of O. trifallax.     

阔口尖毛虫形成包囊期间细胞超微结构的观察

阔口尖毛虫形成囊期间,细胞质内出现条带状或管产产的内质网和由不同大小的囊泡组成的包囊壁前体。并且,前体的产生与内质网有关;细胞质内发生自噬泡消化现象,这是细胞将原有结构和能量进行贮存,利用的一种重要形式;大核向细胞质突出形成阿米巴形结构,这与大核向细胞质排出部分核物质有关。     

阔口尖毛虫无性生殖和生理改组过程的比较

阔口尖毛虫无性生殖中先后发生后口围带原基,前、后的波动膜原基,额腹横棘毛原基和左、右缘棘毛原基,老口围带也在此期间更新,结果形成2套新纤毛结构,原老结构瓦解消失;生理改组时按同样顺序产生口围带原基等几类腹面纤毛原基,结果形成1套新纤毛结构,替换老结构。在这两个截然不同的过程中,新纤毛结构的分化和老纤毛结构退化时也表现出某些相似的特征。作者据此推测,这种纤毛虫无性生殖和生理改组中,纤毛原基的发生、发育和定位在细胞控制机理上可能是相同的。     

Reversible carnivory in Oxytricha bifaria: a peculiar ecological adaptation

The adaptive significance of the giant form of Oxytricha bifaria has been studied. It proved to be a carnivorous phase of a bacterivorous ciliate, rather than a cannibalistic one. By such a dramatic reversal in the feeding behavior O. bifaria, a primary consumer, can reversibly become a secondary one.