Multiple forms of tubulin in the cilia and cytoplasm of Tetrahymena thermophila.

Suprenant, Kathy A.; Hays, Eva; LeCluyse, Edward L.; Dentler, William L.

doi:10.1073/pnas.82.20.6908

Cited by 44 publications

(35 citation statements)

References 63 publications

(53 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunoblot analysis was used to verify the specificity of this antibody. The migration profiles of Tetrahymena ␣-tubulin and ␤-tubulin on SDS-PAGE are considerably different from those of mammalian or avian tubulin; mobility of ␣-tubulin in Tetrahymena is faster than that of ␤-tubulin [Suprenant et al, 1985]. The monoclonal antibody against chick ␣-tubulin recognized a single band on a Western blot containing total or ciliary proteins of T. pyriformis, corresponding to ␣-tubulin (Fig.…”

Section: Intramacronuclear Microtubules Undergo Dramatic Reorganizatimentioning

confidence: 96%

Reorganization of microtubules in the amitotically dividing macronucleus ofTetrahymena

Fujiu

Numata

2000

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

Section: Intramacronuclear Microtubules Undergo Dramatic Reorganizatimentioning

confidence: 96%

Reorganization of microtubules in the amitotically dividing macronucleus ofTetrahymena

Fujiu

Numata

2000

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

“…1). Note that, under conditions described under "Materials and Methods," ciliate ␣-tubulins migrate faster than ␤-tubulins (24). Noticeably, a striking difference in reactivity with the anti-glutamylated tubulin mAb, GT335, indicated the presence of a higher level of glutamylation in Tetrahymena than in Paramecium (Fig.…”

Section: Post-translational Modifications Of Axonemal Tubulins Ofmentioning

confidence: 99%

“…In these highly differentiated cells, the microtubular networks are involved in nuclear divisions, intracellular transport, organelle positioning, and are associated with specialized organelles that function in osmotic regulation, feeding, excretion, and locomotion. The ␣-and ␤-tubulins of ciliates are biochemically heterogeneous (17,20,24), suggesting that structural differences among tubulin isoforms are important in generating functionally distinct types of microtubules in a single cell. However, genetic and biochemical studies showed that in ciliates only one or two gene isotypes of ␣-and ␤-tubulin form the bulk of microtubules (6,(25)(26)(27)(28).…”

mentioning

confidence: 99%

Mutations of Tubulin Glycylation Sites Reveal Cross-talk between the C Termini of α- and β-Tubulin and Affect the Ciliary Matrix in Tetrahymena

Redeker

Levilliers

Vinolo

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Two types of polymeric post-translational modifications of ␣/␤-tubulin, glycylation and glutamylation, occur widely in cilia and flagella. Their respective cellular functions are poorly understood. Mass spectrometry and immunoblotting showed that two closely related species, the ciliates Tetrahymena and Paramecium, have dramatically different compositions of tubulin post-translational modifications in structurally identical axonemes. Whereas the axonemal tubulin of Paramecium is highly glycylated and has a very low glutamylation content, the axonemal tubulin of Tetrahymena is glycylated and extensively glutamylated. In addition, only the ␣-tubulin of Tetrahymena undergoes detyrosination. Mutations of the known glycylation sites in Tetrahymena tubulin affected the level of each polymeric modification type in both the mutated and nonmutated subunits, revealing cross-talk between ␣-and ␤-tubulin. Ultrastructural analyses of glycylation site mutants uncovered defects in the doublet B-subfiber of axonemes and revealed an accumulation of dense material in the ciliary matrix, reminiscent of intraflagellar transport particles seen by others in Chlamydomonas. We propose that polyglycylation and/or polyglutamylation stabilize the B-subfiber of outer doublets and regulate the intraflagellar transport.Microtubules are subject to a set of post-translational modifications (PTMs) 1 whose significance has emerged only recently (1-4). Among PTMs, two polymeric modifications, glutamylation and glycylation, substantially increase the heterogeneity of the ␣/␤-tubulin heterodimer. These tubulin modifications, referred to as polyglutamylation and polyglycylation, correspond to the addition of a peptide polymer consisting of several glutamates (5) or glycines (6) onto the ␥-carboxyl group of a glutamate of the primary sequence of tubulin. These two PTMs will be referred to as "polymodifications" throughout this report. Polymodifications generate peptide branches of variable lengths distributed on several glutamate acceptor sites in the C-terminal tails of ␣-and ␤-tubulin (7-9). Both polymodification types are enriched in flagella and cilia of protists and metazoan cells (4) and were implicated in axoneme motility (10, 11). Whereas polyglycylation is restricted to axonemes in the ciliated and flagellated metazoan cells, polyglutamylation occurs in both axonemes and basal bodies (12-15). In ciliates, both polymodification types are not only present in cilia and basal bodies (15-18), but also occur on the more dynamic intracytoplasmic microtubules (17,19,20). Ciliates assemble up to 17 types of distinct microtubular arrays in a single cell (19,(21)(22)(23). In these highly differentiated cells, the microtubular networks are involved in nuclear divisions, intracellular transport, organelle positioning, and are associated with specialized organelles that function in osmotic regulation, feeding, excretion, and locomotion. The ␣-and ␤-tubulins of ciliates are biochemically heterogeneous (17,20,24), suggesting that structural differences amon...

show abstract

“…Proteins were separated by SDS-PAGE on 10% polyacrylamide minigels (Laemmli, 1970), containing 0.1% (wt/wt) SDS (99% pure; BDH, Poole, United Kingdom) at pH 8.3, according to Suprenant et al (1985). Under these conditions, Paramecium ␣-tubulin migrates faster than ␤-tubulin.…”

Section: Gel Electrophoresis and Immunoblottingmentioning

confidence: 99%

Tubulin Polyglycylation: Differential Posttranslational Modification of Dynamic Cytoplasmic and Stable Axonemal Microtubules inParamecium

Bré

Redeker

Vinh

et al. 1998

MBoC

View full text Add to dashboard Cite

Polyglycylation, a posttranslational modification of tubulin, was discovered in the highly stable axonemal microtubules of Paramecium cilia where it involves the lateral linkage of up to 34 glycine units per tubulin subunit. The observation of this type of posttranslational modification mainly in axonemes raises the question as to its relationship with axonemal organization and with microtubule stability. This led us to investigate the glycylation status of cytoplasmic microtubules that correspond to the dynamic microtubules in Paramecium. Two anti-glycylated tubulin monoclonal antibodies (mAbs), TAP 952 and AXO 49, are shown here to exhibit different affinities toward mono-and polyglycylated synthetic tubulin peptides. Using immunoblotting and mass spectrometry, we show that cytoplasmic tubulin is glycylated. In contrast to the highly glycylated axonemal tubulin, which is recognized by the two mAbs, cytoplasmic tubulin reacts exclusively with TAP 952, and the ␣-and ␤-tubulin subunits are modified by only 1-5 and 2-9 glycine units, respectively. Our analyses suggest that most of the cytoplasmic tubulin contains side chain lengths of 1 or 2 glycine units distributed on several glycylation sites. The subcellular partition of distinct polyglycylated tubulin isoforms between cytoplasmic and axonemal compartments implies the existence of regulatory mechanisms for glycylation. By following axonemal tubulin immunoreactivity with anti-glycylated tubulin mAbs upon incubation with a Paramecium cellular extract, the presence of a deglycylation enzyme is revealed in the cytoplasm of this organism. These observations establish that polyglycylation is reversible and indicate that, in vivo, an equilibrium between glycylating and deglycylating enzymes might be responsible for the length of the oligoglycine side chains of tubulin.

show abstract

Multiple forms of tubulin in the cilia and cytoplasm of Tetrahymena thermophila.

Cited by 44 publications

References 63 publications

Reorganization of microtubules in the amitotically dividing macronucleus ofTetrahymena

Reorganization of microtubules in the amitotically dividing macronucleus ofTetrahymena

Mutations of Tubulin Glycylation Sites Reveal Cross-talk between the C Termini of α- and β-Tubulin and Affect the Ciliary Matrix in Tetrahymena

Tubulin Polyglycylation: Differential Posttranslational Modification of Dynamic Cytoplasmic and Stable Axonemal Microtubules inParamecium

Contact Info

Product

Resources

About