Organization of microtubules in stabilized meristematic plant cells revealed by a rat monoclonal antibody reacting only with the tyrosinated form of ?-tubulin

Wehland, Jürgen; Schroeder, Martin; Weber, Klaus

doi:10.1016/0309-1651(84)90081-x

Cited by 16 publications

(3 citation statements)

References 12 publications

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“…For example, the presence of g-tubulin isoforms correlates with MT stabil-ity (for reviews see [9,22]). Basic information has been improved by use of specific antibodies to different tubufin isoforms in both animal [13,19,21] and plant cells [2,12,26].…”

Section: Discussionmentioning

confidence: 99%

“…The plant microtubular cytoskeleton also consists of different tubulin isoforms. There have been reports on the presence of tyrosinated ~-tubulin in meristematic root cells of Allium [26], the presence of the acetylated form of ~-tubulin in interphase MTs, preprophase band and phragmoplast from root tips of Secale cereale [12] and different tubulin isoforms in pollen tubes grown in agar medium [2].…”

Section: Discussionmentioning

confidence: 99%

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Distribution of microtubules during the growth of tobacco pollen tubes

Del-Casino

1993

Biology of the Cell

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The distribution of microtubules was investigated in Nicotiana tabacum pollen tubes at different stages of tube growth by immunofluorescence microscopy. Using specific antibodies, the presence of microtubules consisting of different tubulin isoforms was tested, a-,/3-and tyrosinated ~t-tubulin were present within the tube, whereas the acetylated form was lacking. The presence of tubulin subunits in pollen tube extracts was al~o investigated by immunobiotting analyses. The use of a confocal laser scanning microscope integrated with computer-assisted imaging, allowed a detailed visualization of the microtubule distribution and organization. Cytoplasmic microtubules organized as short bundles with various orientations were detected at the apex of long tubes. Nicotiana tabacum I pollen tube growth / microtubules / tyrosinated ~-tubulin / acetylated ct-tubulin / confocal laser scanning microscope

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Distribution of microtubules during the growth of tobacco pollen tubes

Del-Casino

1993

Biology of the Cell

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“…In agreement, the monoclonal antibody YL 1/2 (15), which recognizes only the carboxyl end of tyrosinated a-tubulin (40,42), has a broad cross-species reactivity extending from yeast to man and higher plants (15, 40--42). Removal of the tyrosine abolishes antibody reactivity (40,41). Various experiments including some with YL 1/2 show that the carboxy-terminal domains of a-and 13-tubulin are not directly involved in the formation of the microtubular structure.…”

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confidence: 99%

Tubulin-tyrosine ligase has a binding site on beta-tubulin: a two-domain structure of the enzyme

Wehland¹,

Weber²

1987

The Journal of Cell Biology

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Abstract. Tubulin-tyrosine ligase and a~-tubulin form a tight complex which is conveniently monitored by glycerol gradient centrifugation. Using two distinct ligase monoclonal antibodies, several subunit-specific tubulin monoclonal antibodies, and chemical crosslinking, a ligase-binding site was identified on 13-tubulin. This site is retained when the carboxyterminal domains of both tubulin subunits are removed by subtilisin treatment. The ligase-tubulin complex is also formed when ligase is added to al3-tubulin carrying the monoclonal antibody YL 1/2 which binds only to the carboxyl end of tyrosinated a-tubulin. The 13-tubulin-binding site described here explains the extreme substrate specificity of ligase, which does not act on other cellular proteins or carboxy-terminal peptides derived from detyrosinated a-tubulin. Differential accessibility of this site in tubulin and in microtubules seems to explain why ligase acts preferentially on unpolymerized tubulin. Ligase exposed to VS-protease is converted to a nicked derivative. This is devoid of enzymatic activity but still forms the complex with tubulin. Gel electrophoresis documents both 30-and a 14-kD domains, each which is immunologically and biochemically distinct and seems to cover the entire molecule. The two domains interact tightly under physiological conditions. The 30-kD domain carries the binding sites for 13-tubulin and ATP. The 14-kD domain can possibly form an additional part of the catalytic site as it harbors the epitope for the monoclonal antibody ID3 which inhibits enzymatic activity but not the formation of the ligase-tubulin complex.THOUGH encoded by the mRNA (38), the carboxy-terminal tyrosine of a-tubulin shows turnover in cells and tissues (1,4,5,(22)(23)(24). While the process of detyrosination by a putative carboxypeptidase is still poorly understood (2, 3, 17), certain aspects of the reverse reaction are well established in vitro (4,5,(23)(24)(25). Tubulin-tyrosine ligase recharges the detyrosinated a-tubulin of al3-tubulin in an ATP-dependent reaction (5, 24). The enzyme is highly specific for a-tubulin and does not act on other proteins present in the cell or in crude extracts (1, 4, 23). The reaction seems to occur on soluble al3-tubulin rather than on microtubules (1, 23, 36) in line with an isolatable complex formed between ligase and al3-tubulin (20,24,28). Ligase activity has been found in various vertebrates (22, 34), in several invertebrates (10, 16), and even in protozoa (32). In addition, all but two of the many a-tubulin genes described to date predict the presence of a carboxy-terminal tyrosine residue (7,14,39). In agreement, the monoclonal antibody YL 1/2 (15), which recognizes only the carboxyl end of tyrosinated a-tubulin (40, 42), has a broad cross-species reactivity extending from yeast to man and higher plants (15, 40--42). Removal of the tyrosine abolishes antibody reactivity (40,41). Various experiments including some with YL 1/2 show that the carboxy-terminal domains of a-and 13-tubulin are not directly involved i...

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Man toTrypanosome: The tubulin tyrosination/detyrosination cycle revisited

Idriss

2000

Cell Motil. Cytoskeleton

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Organization of microtubules in stabilized meristematic plant cells revealed by a rat monoclonal antibody reacting only with the tyrosinated form of ?-tubulin

Cited by 16 publications

References 12 publications

Distribution of microtubules during the growth of tobacco pollen tubes

Distribution of microtubules during the growth of tobacco pollen tubes

Tubulin-tyrosine ligase has a binding site on beta-tubulin: a two-domain structure of the enzyme

Man toTrypanosome: The tubulin tyrosination/detyrosination cycle revisited

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