scholarly journals Patterns of basal body addition in ciliary rows in Tetrahymena.

1975 ◽  
Vol 65 (3) ◽  
pp. 503-512 ◽  
Author(s):  
D L Nanney

Most naked basal bodies visualized in protargol stains on the surface of Tetrahymena are new basal bodies which have not yet developed cilia. The rarity of short cilia is explained by the rapid development of the ciliary shaft once it begins to grow. The high frequency of naked basal bodies (about 50 percent) in log cultures indicates that the interval between assembly of the basal body and the initiation of the cilium is long, approximately a full cell cycle. Naked basal bodies are more frequent in the mid and posterior parts of the cell and two or more naked basal bodies may be associated with one ciliated basal body in these regions. Daughter cells produced at division are apparently asymmetric with respect to their endowment of new and old organelles.

We have analysed the timing and order of events occurring within the cell division cycle of Trypanosoma brucei . Cells in the earliest stages of the cell cycle possess a single copy of three major organelles: the nucleus, the kinetoplast and the flagellum. The first indication of progress through the cell cycle is the elongation of the pro-basal body lying adjacent to the mature basal body subtending the flagellum. This newly elongated basal body occupies a posterior position within the cell when it initiates growth of the new daughter flagellum. Genesis of two new pro-basal bodies occurs only after growth of the new daughter flagellum has been initiated. Extension of the new flagellum, together with the paraflagellar rod, then continues throughout a major portion of the cell cycle. During this period of flagellum elongation, kinetoplast division occurs and the two kinetoplasts, together with the two flagellar basal bodies, then move apart within the cell. Mitosis is then initiated and a complex pattern of organelle positions is achieved whereby a division plane runs longitudinally through the cell such that each daughter ultimately receives a single nucleus, kinetoplast and flagellum. These events have been described from observations of whole cytoskeletons by transmission electron microscopy together with detection of particular organelles by fluorescence microscopy. The order and timing of events within the cell cycle has been derived from analyses of the proportion of a given cell type occurring within an exponentially growing culture.


2008 ◽  
Vol 19 (1) ◽  
pp. 262-273 ◽  
Author(s):  
Brian P. Piasecki ◽  
Matthew LaVoie ◽  
Lai-Wa Tam ◽  
Paul A. Lefebvre ◽  
Carolyn D. Silflow

Mutations in the UNI2 locus in Chlamydomonas reinhardtii result in a “uniflagellar” phenotype in which flagellar assembly occurs preferentially from the older basal body and ultrastructural defects reside in the transition zones. The UNI2 gene encodes a protein of 134 kDa that shares 20.5% homology with a human protein. Immunofluorescence microscopy localized the protein on both basal bodies and probasal bodies. The protein is present as at least two molecular-weight variants that can be converted to a single form with phosphatase treatment. Synthesis of Uni2 protein is induced during cell division cycles; accumulation of the phosphorylated form coincides with assembly of transition zones and flagella at the end of the division cycle. Using the Uni2 protein as a cell cycle marker of basal bodies, we observed migration of basal bodies before flagellar resorption in some cells, indicating that flagellar resorption is not required for mitotic progression. We observed the sequential assembly of new probasal bodies beginning at prophase. The uni2 mutants may be defective in the pathways leading to flagellar assembly and to basal body maturation.


2013 ◽  
Vol 24 (9) ◽  
pp. 1321-1333 ◽  
Author(s):  
Ana Lozano-Núñez ◽  
Kyojiro N. Ikeda ◽  
Thomas Sauer ◽  
Christopher L. de Graffenried

Polo-like kinases are important regulators of cell division, playing diverse roles in mitosis and cytoskeletal inheritance. In the parasite Trypanosoma brucei, the single PLK homologue TbPLK is necessary for the assembly of a series of essential organelles that position and adhere the flagellum to the cell surface. Previous work relied on RNA interference or inhibitors of undefined specificity to inhibit TbPLK, both of which have significant experimental limitations. Here we use an analogue-sensitive approach to selectively and acutely inhibit TbPLK. T. brucei cells expressing only analogue-sensitive TbPLK (TbPLKas) grow normally, but upon treatment with inhibitor develop defects in flagellar attachment and cytokinesis. TbPLK cannot migrate effectively when inhibited and remains trapped in the posterior of the cell throughout the cell cycle. Using synchronized cells, we show that active TbPLK is a direct requirement for the assembly and extension of the flagellum attachment zone, which adheres the flagellum to the cell surface, and for the rotation of the duplicated basal bodies, which positions the new flagellum so that it can extend without impinging on the old flagellum. This approach should be applicable to the many kinases found in the T. brucei genome that lack an ascribed function.


2011 ◽  
Vol 193 (3) ◽  
pp. 435-444 ◽  
Author(s):  
Tetsuo Kobayashi ◽  
Brian D. Dynlacht

The role of centrioles changes as a function of the cell cycle. Centrioles promote formation of spindle poles in mitosis and act as basal bodies to assemble primary cilia in interphase. Stringent regulations govern conversion between these two states. Although the molecular mechanisms have not been fully elucidated, recent findings have begun to shed light on pathways that regulate the conversion of centrioles to basal bodies and vice versa. Emerging studies also provide insights into how defects in the balance between centrosome and cilia function could promote ciliopathies and cancer.


1998 ◽  
Vol 9 (6) ◽  
pp. 1293-1308 ◽  
Author(s):  
Susan K. Dutcher ◽  
Emanuel C. Trabuco

We have cloned the UNI3 gene inChlamydomonas and find that it encodes a new member of the tubulin superfamily. Although Uni3p shares significant sequence identity with α-, β-, and γ-tubulins, there is a region of Uni3p that has no similarity to tubulins or other known proteins. Mutantuni3–1 cells assemble zero, one, or two flagella. Pedigree analysis suggests that flagellar number inuni3–1 cells is a function of the age of the cell. The uniflagellate uni3–1 cells show a positional phenotype; the basal body opposite the eyespot templates the single flagellum. A percentage of uni3–1 cells also fail to orient the cleavage furrow properly, and basal bodies have been implicated in the placement of cleavage furrows in Chlamydomonas. Finally when uni3–1 cells are observed by electron microscopy, doublet rather than triplet microtubules are observed at the proximal end of the basal bodies. We propose that the Uni3 tubulin is involved in both the function and cell cycle-dependent maturation of basal bodies/centrioles.


2009 ◽  
Vol 20 (1) ◽  
pp. 379-388 ◽  
Author(s):  
M. Qasim Rasi ◽  
Jeremy D.K. Parker ◽  
Jessica L. Feldman ◽  
Wallace F. Marshall ◽  
Lynne M. Quarmby

Katanin is a microtubule-severing protein that participates in the regulation of cell cycle progression and in ciliary disassembly, but its precise role is not known for either activity. Our data suggest that in Chlamydomonas, katanin severs doublet microtubules at the proximal end of the flagellar transition zone, allowing disengagement of the basal body from the flagellum before mitosis. Using an RNA interference approach we have discovered that severe knockdown of the p60 subunit of katanin, KAT1, is achieved only in cells that also carry secondary mutations that disrupt ciliogenesis. Importantly, we observed that cells in the process of cell cycle-induced flagellar resorption sever the flagella from the basal bodies before resorption is complete, and we find that this process is defective in KAT1 knockdown cells.


1999 ◽  
Vol 112 (24) ◽  
pp. 4641-4650 ◽  
Author(s):  
A. Ploubidou ◽  
D.R. Robinson ◽  
R.C. Docherty ◽  
E.O. Ogbadoyi ◽  
K. Gull

Trypanosoma brucei has a single nucleus and a single kinetoplast (the mitochondrial genome). Each of these organelles has a distinct S phase, which is followed by a segregation period, prior to cell division. The segregation of the two genomes takes place in a specific temporal order by interaction with microtubule-based structures, the spindle for nuclear DNA and the flagellum basal bodies for the kinetoplast DNA. We used rhizoxin, the anti-microtubule agent and polymerisation inhibitor, or the nuclear DNA synthesis inhibitor aphidicolin, to interfere with cell cycle events in order to study how such events are co-ordinated. We show that T. brucei cytokinesis is not dependent upon either mitosis or nuclear DNA synthesis, suggesting that there are novel cell cycle checkpoints in this organism. Moreover, use of monoclonal antibodies to reveal cytoplasmic events such as basal body duplication shows that some aphidicolin treated cells appear to be in G(1) phase (1K1N) but have activated some cytoplasmic events characteristic of G(2) phase (basal body segregation). We discuss a possible dominant role in trypanosomes for kinetoplast/basal body segregation in control of later cell cycle events such as cytokinesis


PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0249908
Author(s):  
Catherine Sullenberger ◽  
Benjamin Hoffman ◽  
Justin Wiedeman ◽  
Gaurav Kumar ◽  
Kojo Mensa-Wilmot

The single mitochondrial nucleoid (kinetoplast) of Trypanosoma brucei is found proximal to a basal body (mature (mBB)/probasal body (pBB) pair). Kinetoplast inheritance requires synthesis of, and scission of kinetoplast DNA (kDNA) generating two kinetoplasts that segregate with basal bodies into daughter cells. Molecular details of kinetoplast scission and the extent to which basal body separation influences the process are unavailable. To address this topic, we followed basal body movements in bloodstream trypanosomes following depletion of protein kinase TbCK1.2 which promotes kinetoplast division. In control cells we found that pBBs are positioned 0.4 um from mBBs in G1, and they mature after separating from mBBs by at least 0.8 um: mBB separation reaches ~2.2 um. These data indicate that current models of basal body biogenesis in which pBBs mature in close proximity to mBBs may need to be revisited. Knockdown of TbCK1.2 produced trypanosomes containing one kinetoplast and two nuclei (1K2N), increased the percentage of cells with uncleaved kDNA 400%, decreased mBB spacing by 15%, and inhibited cytokinesis 300%. We conclude that (a) separation of mBBs beyond a threshold of 1.8 um correlates with division of kDNA, and (b) TbCK1.2 regulates kDNA scission. We propose a Kinetoplast Division Factor hypothesis that integrates these data into a pathway for biogenesis of two daughter mitochondrial nucleoids.


Development ◽  
1990 ◽  
Vol 109 (4) ◽  
pp. 935-942 ◽  
Author(s):  
N.E. Williams ◽  
J.E. Honts ◽  
J. Kaczanowska

Differentiated regions within the membrane skeleton are described around basal bodies in the ciliary rows of Tetrahymena. These domains, approximately 1 micron in diameter, are characterized by a relatively dense ultrastructure, the presence of a family of proteins called K antigens (Mr 39–44 × 10(3)) that are recognized by mAb 424A8, and the apparent exclusion of major membrane skeleton proteins that are present in most other regions of the cell (Mr 135, 125 × 10(3]. Mature basal body domains are asymmetric, reflecting the polarity of the cell as a whole. A similar differentiation of the membrane skeleton occurs in the oral apparatus, except here the K antigens surround four clusters of basal bodies (from which this cell takes its name) rather than the individual basal bodies. The development of new basal body domains in the cell cycle is described, with similarities and differences noted between somatic and oral regions of the cell. It is concluded that the capacity of this cell for precise topographic regulation of molecular events in the membrane skeleton makes it a useful model for the study of cortical differentiation in cells generally.


1970 ◽  
Vol 7 (1) ◽  
pp. 65-89
Author(s):  
M. FRIEDLÄNDER ◽  
J. WAHRMAN

The spindle of metazoan cells functions as a dual distributor that guarantees the accurate segregation of both chromosomes and centrioles (basal bodies). This combined mechanism may have evolved from the separate distribution devices for chromosomes and basal bodies found in Protozoa. Typical eupyrene spermatocytes of the silk moth were compared with atypical apyrene spermatocytes. (a) Long microtubules, persisting during centriolar movements, develop in the meiotic prophases in continuity with the centrioles (b) There is no longer a centriole-microtubule continuity at metaphase-anaphase (c) Spindles comprise microtubules and endoplasmic reticulum (ER). The microtubules form a barrel-shaped structure terminating far in front of the centrioles. The two types of spermatocytes differ in the structure of the ER, which is vesicular in the eupyrene and lamellar in the apyrene line. It is suggested that the ER influences the anaphase movement of chromosomes, (d) The chromosomes lack localized centromeres. Microtubules penetrate all along the polar faces of the eupyrene metaphase chromosomes. The apyrene chromosomes form irregular blocks that are penetrated by microtubules all around their periphery, (e) The centriole comprises 3 concentric zones. Typical changes in the centriole are correlated with changes in the cell cycle. Replication, de novo formation and disappearance of centrioles are manifestations of the state of flux of the microtubules.


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