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1.
Antisera against electrophoretically purified tubulin stimulate colchicine-binding activity. 总被引:2,自引:0,他引:2
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J E Aubin L Subrahmanyan V I Kalnins V Ling 《Proceedings of the National Academy of Sciences of the United States of America》1976,73(4):1246-1249
Several rabbit antisera have been prepared against reduced and alkylated, electrophoretically purified tubulin isolated from chick brain. These antisera give a single precipitin line in Ouchterlony double diffusion plates when tested against partially purified tubulin, and label specifically microtubule- and tubulin-containing structures, such as mitotic spindles, cilia, and vinblastine-induced crystals, in a variety of cells. The same antisera also display the unique ability to stimulate the colchicine-binding activity of tubulin preparations from chick brain and Chinese hamster ovary tissue culture cells. This specific stimulation of colchicine binding activity is also obtained with the gamma globulin fractions purified by ammonium sulfate precipitation of these antisera. 相似文献
2.
ASAP, a human microtubule-associated protein required for bipolar spindle assembly and cytokinesis
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Saffin JM Venoux M Prigent C Espeut J Poulat F Giorgi D Abrieu A Rouquier S 《Proceedings of the National Academy of Sciences of the United States of America》2005,102(32):11302-11307
We have identified a unique human microtubule-associated protein (MAP) named ASAP for ASter-Associated Protein. ASAP localizes to microtubules in interphase, associates with the mitotic spindle during mitosis, localizes to the central body during cytokinesis and directly binds to purified microtubules by its COOH-terminal domain. Overexpression of ASAP induces profound bundling of cytoplasmic microtubules in interphase cells and aberrant monopolar spindles in mitosis. Depletion of ASAP by RNA interference results in severe mitotic defects: it provokes aberrant mitotic spindle, delays mitotic progression, and leads to defective cytokinesis or cell death. These results suggest a crucial role for ASAP in the organization of the bipolar mitotic spindle, mitosis progression, and cytokinesis and define ASAP as a key factor for proper spindle assembly. 相似文献
3.
Benjamin R. Houghtaling Ge Yang Alexandre Matov Gaudenz Danuser Tarun M. Kapoor 《Proceedings of the National Academy of Sciences of the United States of America》2009,106(36):15338-15343
Accuracy in chromosome segregation depends on the assembly of a bipolar spindle. Unlike mitotic spindles, which have roughly equal amounts of kinetochore microtubules (kMTs) and nonkinetochore microtubules (non-kMTs), vertebrate meiotic spindles are predominantly comprised of non-kMTs, a large subset of which forms an antiparallel “barrel” array at the spindle equator. Though kMTs are needed to drive chromosome segregation, the contributions of non-kMTs are more mysterious. Here, we show that increasing the concentration of Op18/stathmin, a component of the chromosome-mediated microtubule formation pathway that directly controls microtubule dynamics, can be used to deplete non-kMTs in the vertebrate meiotic spindle assembled in Xenopus egg extracts. Under these conditions, kMTs and the spindle pole-associated non-kMT arrays persist in smaller spindles. In excess Op18, distances between sister kinetochores, an indicator of tension across centromeres, remain unchanged, even though kMTs flux poleward with a ≈30% slower velocity, and chromosomes oscillate more than in control metaphase spindles. Remarkably, kinesin-5, a conserved motor protein that can push microtubules apart and is required for the assembly and maintenance of bipolar meiotic spindles, is not needed to maintain spindle bipolarity in the presence of excess Op18. Our data suggest that non-kMTs in meiotic spindles contribute to normal kMT dynamics, stable chromosome positioning, and the establishment of proper spindle size. We propose that without non-kMTs, metaphase meiotic spindles are similar to mammalian mitotic spindles, which balance forces to maintain metaphase spindle organization in the absence of extensive antiparallel microtubule overlap at the spindle equator or a key mitotic kinesin. 相似文献
4.
G J Gorbsky C Simerly G Schatten G G Borisy 《Proceedings of the National Academy of Sciences of the United States of America》1990,87(16):6049-6053
After ovulation mammalian oocytes arrest in second meiotic metaphase. We asked whether the microtubules that comprise the meiotic spindle of mouse oocytes were stable or were undergoing rapid cycles of assembly and disassembly. Porcine brain tubulin, derivatized with biotin or x-rhodamine [5- (and -6)-carboxy-x-rhodamine], was microinjected into living oocytes. Biotinylated tubulin incorporated into the meiotic spindle to apparent equilibrium within 15 min. To assess quantitatively the rates of disassembly and assembly of the microtubules, small domains within the spindles of oocytes injected with x-rhodamine-tubulin were photobleached and their recovery was analyzed by digital imaging microscopy. Fluorescence recovery in the spindles was rapid and extensive, plateauing to an average of 83% at 4 min. The calculated half-time for turnover of the spindle microtubules was 77 sec. In contrast, fluorescence recovery of the spindle midbodies in telophase oocytes was much more limited, averaging approximately 22% at 4 min. These data indicate that most microtubules within the arrested metaphase spindle of the mouse oocyte undergo rapid cycles of assembly and disassembly. Microtubules of the telophase midbody are more stable. 相似文献
5.
M D Weingarten A H Lockwood S Y Hwo M W Kirschner 《Proceedings of the National Academy of Sciences of the United States of America》1975,72(5):1858-1862
A heat stable protein essentail for microtubule assembly has been isolated. This protein, which we designate tau (tau), is present in association with tubulin purified from porcine brain by repeated cycles of polymerization. Tau is separated from tubulin by ion exchange chromatography on phosphocellulose. In the absence of tau, tubulin exists entirely as a 6S dimer of two polypeptide chains (alpha and beta tubulin) with a molecular weight of 120,000, which will not assemble into microtubules in vitro. Addition of tau completely restores tubule-forming capacity. Under nonpolymerizing conditions, tau converts 6S dimers to 36S rings-structures which have been implicated as intermediates in tubule formation. Hence, tau appears to act on the 6S tubulin dimer, activating it for polymerization. The unique ability of tau to restore the normal features of in vitro microtubule assembly makes it likely that tau is a major regulator of microtubule formation in cells. 相似文献
6.
Evidence for two distinct binding sites for tau on microtubules 总被引:1,自引:0,他引:1
Makrides V Massie MR Feinstein SC Lew J 《Proceedings of the National Academy of Sciences of the United States of America》2004,101(17):6746-6751
The microtubule-associated protein tau regulates diverse and essential microtubule functions, from the nucleation and promotion of microtubule polymerization to the regulation of microtubule polarity and dynamics, as well as the spacing and bundling of axonal microtubules. Thermodynamic studies show that tau interacts with microtubules in the low- to mid-nanomolar range, implying moderate binding affinity. At the same time, it is well established that microtubule-bound tau does not undergo exchange with the bulk medium readily, suggesting that the tau-microtubule interaction is essentially irreversible. Given this dilemma, we investigated the mechanism of interaction between tau and microtubules in kinetic detail. Stopped-flow kinetic analysis reveals moderate binding affinity between tau and preassembled microtubules and rapid dissociation/association kinetics. In contrast, when microtubules are generated by copolymerization of tubulin and tau, a distinct population of microtubule-bound tau is observed, the binding of which seems irreversible. We propose that reversible binding occurs between tau and the surface of preassembled microtubules, whereas irreversible binding results when tau is coassembled with tubulin into a tau-microtubule copolymer. Because the latter is expected to be physiologically relevant, its characterization is of central importance. 相似文献
7.
Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. 总被引:17,自引:2,他引:17
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G Schatten C Simerly H Schatten 《Proceedings of the National Academy of Sciences of the United States of America》1985,82(12):4152-4156
Microtubules forming within the mouse egg during fertilization are required for the movements leading to the union of the sperm and egg nuclei (male and female pronuclei, respectively). In the unfertilized oocyte, microtubules are predominantly found in the arrested meiotic spindle. At the time for sperm incorporation, a dozen cytoplasmic asters assemble, often associated with the pronuclei. As the pronuclei move to the egg center, these asters enlarge into a dense array. At the end of first interphase, the dense array disassembles and is replaced by sheaths of microtubules surrounding the adjacent pronuclei. Syngamy (pronuclear fusion) is not observed; rather the adjacent paternal and maternal chromosome sets first meet at metaphase. The mitotic apparatus emerges from these perinuclear microtubules and is barrel-shaped and anastral, reminiscent of plant cell spindles; the sperm centriole does not nucleate mitotic microtubules. After cleavage, monasters extend from each blastomere nucleus. The second division mitotic spindles also have broad poles, though by third and later divisions the spindles are typical for higher animals, with narrow mitotic poles and fusiform shapes. Colcemid, griseofulvin, and nocodazole inhibit the microtubule formation and prevent the movements leading to pronuclear union; the meiotic spindle is disassembled, and the maternal chromosomes are scattered throughout the oocyte cortex. These results indicate that microtubules forming within fertilized mouse oocytes are required for the union of the sperm and egg nuclei and raise questions about the paternal inheritance of centrioles in mammals. 相似文献
8.
Isolation of sea urchin egg microtubules with taxol and identification of mitotic spindle microtubule-associated proteins with monoclonal antibodies. 总被引:18,自引:3,他引:18
R B Vallee G S Bloom 《Proceedings of the National Academy of Sciences of the United States of America》1983,80(20):6259-6263
Microtubules were isolated from unfertilized eggs of the sea urchin with the use of the anti-tumor drug taxol. In addition to tubulin, prominent high molecular weight (Mr 205,000-350,000) microtubule-associated proteins (MAPs) were identified as well as MAP species of Mrs 77,000, 100,000, and 120,000. The microtubules were covered with both short periodic arms and longer filamentous arms, both classes of which appeared to crosslink the microtubules into bundles. Monoclonal antibodies were prepared to an unfractionated MAPs preparation. We isolated clonal hybridoma lines producing antibodies to tubulin and to four non-tubulin proteins of Mrs 235,000, 205,000, 150,000, and 37,000. All antibodies strongly and specifically stained the mitotic spindle of dividing sea urchin eggs. All four of the immunoreactive, non-tubulin species behaved as MAPs during microtubule isolation. Thus, we have identified a variety of sea urchin MAPs by biochemical, ultrastructural, and immunochemical means. The immunochemical experiments demonstrated that four of these proteins are microtubule-associated components of the mitotic spindle. We suggest that those proteins that we observed as cross-bridges between the isolated microtublules may be either structural or functional components of the spindle. 相似文献
9.
G B Witman D W Cleveland M D Weingarten M W Kirschner 《Proceedings of the National Academy of Sciences of the United States of America》1976,73(11):4070-4074
Tubulin purified by phosphocellulose chromatography and free of accessory proteins will not form microtubules in the absence or presence of microtubule initiating sites (flagellar microtubules). The capacity for growth onto pre-existing "seeds" can be restored by the addition of small quantities of partially purified tau protein. Larger quantities restore the capacity for spontaneous assembly. These results suggest that tubulin requires tau for both initiation and growth of microtubules and that tau is incorporated into the microtubule throughout its length. 相似文献
10.
Brain tubulin polymerization in the absence of "microtubule-associated proteins". 总被引:9,自引:0,他引:9
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R H Himes P R Burton R N Kersey G B Pierson 《Proceedings of the National Academy of Sciences of the United States of America》1976,73(12):4397-4399
Pure tubulin dimer, purified by phosphecellulose chromatography to remove microtubule associated proteins, can be assembled into microtubules in the presence of dimethyl sulfoxide. The surfaces of such microtubules are devoid of filamentous material. The reaction is rapid and strongly dependent on protein concentration. No microtubule formation occurs from the purified tubulin when dimethyl sulfoxide is omitted. 相似文献
11.
Comparison of the effects of microtubule-associated protein 2 and tau on the packing density of in vitro assembled microtubules. 总被引:6,自引:0,他引:6
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M M Black 《Proceedings of the National Academy of Sciences of the United States of America》1987,84(21):7783-7787
I have compared the effects of microtubule-associated protein 2 (MAP-2) and tau on the packing density of sedimented microtubules. Microtubules assembled in vitro in taxol were pelleted by centrifugation. The volumes of the resulting pellets were calculated from their weights assuming a specific gravity of 1 and then were normalized to the amount of protein in the pellet, yielding a value for pellet specific volume in microliter/mg of protein. The specific volume of the pellets reflects the intermicrotubule spacing within the pellet. Microtubules were assembled from tubulin alone or tubulin plus various amounts of MAP-2 or tau and collected by centrifugation, and the pellet specific volume was measured. The specific volume of microtubules composed of pure tubulin ranged from 6.4 to 7.7 microliter/mg of protein. Tau had no detectable effect on this value even at saturating levels on the microtubules. In contrast, MAP-2 increased pellet specific volume as the MAP-2/tubulin weight ratio increased; at the highest ratio examined, 0.43, the pellet specific volume was approximately 33. Even at the relatively low MAP-2/tubulin ratio of 0.09, pellet specific volume was approximately 2-fold greater than that of microtubules containing tubulin alone or tubulin plus tau. Electron microscopy confirmed that the observations on pellet specific volume reflected differences in the effects of MAP-2 and tau on the packing density of sedimented microtubules. These results are discussed in the context of observations showing that neighboring microtubules are more widely spaced in dendrites than in axons and that MAP-2 is enriched on microtubules in dendrites compared to microtubules in axons, whereas the converse is true for tau. 相似文献
12.
Kolano A Brunet S Silk AD Cleveland DW Verlhac MH 《Proceedings of the National Academy of Sciences of the United States of America》2012,109(27):E1858-E1867
It is well established that chromosome segregation in female meiosis I (MI) is error-prone. The acentrosomal meiotic spindle poles do not have centrioles and are not anchored to the cortex via astral microtubules. By Cre recombinase-mediated removal in oocytes of the microtubule binding site of nuclear mitotic apparatus protein (NuMA), which is implicated in anchoring microtubules at poles, we determine that without functional NuMA, microtubules lose connection to MI spindle poles, resulting in highly disorganized early spindle assembly. Subsequently, very long spindles form with hyperfocused poles. The kinetochores of homologs make attachments to microtubules in these spindles but with reduced tension between them and accompanied by alignment defects. Despite this, the spindle assembly checkpoint is normally silenced and the advance to anaphase I and first polar body extrusion takes place without delay. Females without functional NuMA in oocytes are sterile, producing aneuploid eggs with altered chromosome number. These findings establish that in mammalian MI, the spindle assembly checkpoint is unable to sustain meiotic arrest in the presence of one or few misaligned and/or misattached kinetochores with reduced interkinetochore tension, thereby offering an explanation for why MI in mammals is so error-prone. 相似文献
13.
Role of abnormally phosphorylated tau in the breakdown of microtubules in Alzheimer disease. 总被引:15,自引:0,他引:15
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A C Alonso T Zaidi I Grundke-Iqbal K Iqbal 《Proceedings of the National Academy of Sciences of the United States of America》1994,91(12):5562-5566
The microtubule assembly-promoting activity of different pools of tau protein isolated from Alzheimer disease (AD) and control brains and the effect of dephosphorylation on this activity were studied. Tau isolated from a 2.5% perchloric extract of AD brain had almost the same activity as that obtained from control brain, and this activity did not change significantly on dephosphorylation. Abnormally phosphorylated tau (AD P-tau) isolated from brain homogenate of AD patients had little activity, and upon dephosphorylation with alkaline phosphatase, its activity increased to approximately the same level as the acid-soluble tau. Addition of AD P-tau to a mixture of normal tau and tubulin inhibited microtubule assembly. AD P-tau bound to normal tau but not to tubulin. These studies suggest that the abnormal phosphorylation of tau might be responsible for the breakdown of microtubules in affected neurons in AD not only because the altered protein has little microtubule-promoting activity but also because it interacts with normal tau, making the latter unavailable for promoting the assembly of tubulin into microtubules. 相似文献
14.
Drosophila Mgr, a Prefoldin subunit cooperating with von Hippel Lindau to regulate tubulin stability
Delgehyr N Wieland U Rangone H Pinson X Mao G Dzhindzhev NS McLean D Riparbelli MG Llamazares S Callaini G Gonzalez C Glover DM 《Proceedings of the National Academy of Sciences of the United States of America》2012,109(15):5729-5734
Mutations in Drosophila merry-go-round (mgr) have been known for over two decades to lead to circular mitotic figures and loss of meiotic spindle integrity. However, the identity of its gene product has remained undiscovered. We now show that mgr encodes the Prefoldin subunit counterpart of human von Hippel Lindau binding-protein 1. Depletion of Mgr from cultured cells also leads to formation of monopolar and abnormal spindles and centrosome loss. These phenotypes are associated with reductions of tubulin levels in both mgr flies and mgr RNAi-treated cultured cells. Moreover, mgr spindle defects can be phenocopied by depleting β-tubulin, suggesting Mgr function is required for tubulin stability. Instability of β-tubulin in the mgr larval brain is less pronounced than in either mgr testes or in cultured cells. However, expression of transgenic β-tubulin in the larval brain leads to increased tubulin instability, indicating that Prefoldin might only be required when tubulins are synthesized at high levels. Mgr interacts with Drosophila von Hippel Lindau protein (Vhl). Both proteins interact with unpolymerized tubulins, suggesting they cooperate in regulating tubulin functions. Accordingly, codepletion of Vhl with Mgr gives partial rescue of tubulin instability, monopolar spindle formation, and loss of centrosomes, leading us to propose a requirement for Vhl to promote degradation of incorrectly folded tubulin in the absence of functional Prefoldin. Thus, Vhl may play a pivotal role: promoting microtubule stabilization when tubulins are correctly folded by Prefoldin and tubulin destruction when they are not. 相似文献
15.
A widely distributed nuclear protein immunologically related to the microtubule-associated protein MAP1 is associated with the mitotic spindle. 总被引:9,自引:2,他引:7
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J S Bonifacino R D Klausner I V Sandoval 《Proceedings of the National Academy of Sciences of the United States of America》1985,82(4):1146-1150
A 280-kDa protein (p280) confined to the nucleus of interphase cells becomes associated with the mitotic spindle during cell division. p280 is immunologically related to the microtubule-associated protein MAP1, as shown by cross-reactivity with monoclonal (8D12) and polyclonal antibodies raised against MAP1. However, p280 is distinct from MAP1 as judged by its lower molecular size, proteolytic degradation products, presence in preparations of purified nuclei from which MAP1 is absent, and absence from the cytosol fraction that contains MAP1. Immunofluorescence microscopy of cells in interphase using 8D12 reveals punctate staining of the nucleus, cytoplasmic microtubules, and the microtubule organizing center. Dividing cells display strong staining of the spindle, centrioles, and mid-body. The only exception to this staining pattern is marsupial Pt k2 cells that contain p280 in the nucleus and lack MAP1. These cells exhibit fluorescent staining of the nucleus and the microtubule organizing center when in interphase, of spindle and centrioles in mitosis, and show no staining of cytoplasmic and mid-body microtubules. 相似文献
16.
Kinesin is associated with a nonmicrotubule component of sea urchin mitotic spindles. 总被引:16,自引:3,他引:13
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R J Leslie R B Hird L Wilson J R McIntosh J M Scholey 《Proceedings of the National Academy of Sciences of the United States of America》1987,84(9):2771-2775
Sea urchin embryos in second division have been lysed into microtubule-stabilizing buffers to yield mitotic cytoskeletons (MCSs) that consist of two mitotic spindles surrounded by a cortical array of filaments. Microtubules have been completely extracted from MCSs by incubation at 0 degrees C with Ca2+-containing buffer. An antibody to the microtubule translocator kinesin stains the spindles in MCSs and in MCSs treated with 5 mM ATP and also stains spindle-remnants of the MCSs after the microtubules have been extracted. We conclude that kinesin binds to a nonmicrotubule component in the mitotic spindle. Based on these results, we present several models of kinesin function in the spindle. 相似文献
17.
Low molecular weight microtubule-associated proteins are light chains of microtubule-associated protein 1 (MAP 1). 总被引:22,自引:7,他引:22
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R B Vallee S E Davis 《Proceedings of the National Academy of Sciences of the United States of America》1983,80(5):1342-1346
Microtubule-associated protein 1 (MAP 1; Mr = 350,000) was analyzed by column chromatography of microtubule protein obtained from calf brain gray and white matter. Two low molecular weight proteins (LMW MAPs; Mr 28,000 and 30,000) were found to cochromatograph with MAP 1 under all conditions examined. MAP 1 and the LMW MAPs were purified from calf brain white matter as a complex containing approximately equimolar amounts of the three species. Urea (6 M) was used to remove the LMW MAPs from MAP 1. Binding of MAP 1 to microtubules was unaffected by urea and occurred with or without the LMW species. Electron microscopy of microtubules composed of purified tubulin and either MAP 1 preparation revealed that, like MAP 2, MAP 1 has the appearance of a filamentous arm on the microtubule surface. 相似文献
18.
Analysis of mitotic microtubule-associated proteins using mass
spectrometry identifies astrin, a spindle-associated protein
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Gary J. Mack Duane A. Compton 《Proceedings of the National Academy of Sciences of the United States of America》2001,98(25):14434-14439
We purified microtubules from a mammalian mitotic extract and obtained an amino acid sequence from each microtubule-associated protein by using mass spectrometry. Most of these proteins are known spindle-associated components with essential functional roles in spindle organization. We generated antibodies against a protein identified in this collection and refer to it as astrin because of its association with astral microtubule arrays assembled in vitro. Astrin is approximately 134 kDa, and except for a large predicted coiled-coil domain in its C-terminal region it lacks any known functional motifs. Astrin associates with spindle microtubules as early as prophase where it concentrates at spindle poles. It localizes throughout the spindle in metaphase and anaphase and associates with midzone microtubules in anaphase and telophase. Astrin also localizes to kinetochores but only on those chromosomes that have congressed. Deletion analysis indicates that astrin's primary spindle-targeting domain is at the C terminus, although a secondary domain in the N terminus can target some of the protein to spindle poles. Thus, we have generated a comprehensive list of major mitotic microtubule-associated proteins, among which is astrin, a nonmotor spindle protein. 相似文献
19.
Double indirect immunofluorescent labeling of embryonic chick tissue was undertaken for the vitamin D-induced calcium binding protein, calbindin-D28k, and microtubules. Immunoreactivities for both calbindin-D28k and tubulin were found to exhibit a filamentous staining pattern in mesonephros, metanephros, intestine, and brain. In the intestine, staining was absent at 19 days, while immunolabeling of tubulin became evident at 20 days, and both antigens were present in 21-day tissue. In intestinal epithelium, as well as in 10-day metanephros, it was strikingly evident that cells either stained for both antigens or were negative for both calbindin-D28k and tubulin. In 11-12-day metanephros, an increased number of cells with both immunoreactivities were found. In 15-17-day brain, tubulin was evident within all cells but stained most intensely in Purkinje cells which were also positive for calbindin-D28k. Mesonephros from 4-5-day embryos revealed immunolabeling of both tubulin and the calcium binding protein. A statistical analysis of the various cell types revealed that the vast majority contained either both antigens or neither of the immunoreactivities. Of the more than 600 cells scored, none were found to be positive for calbindin-D28k, while at the same time negative for tubulin. It is concluded that calbindin-D28k exhibits a noncytoplasmic distribution in all tissues tested and that the filamentous appearance may reflect localization of the antigen in tubulo-vesicular organelles associated with cytoskeleton. 相似文献
20.
Inhibition of tubulin assembly by RNA and other polyanions: evidence for a required protein. 总被引:13,自引:4,他引:9
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J B Bryan B W Nagle K H Doenges 《Proceedings of the National Academy of Sciences of the United States of America》1975,72(9):3570-3574
Nonneural cell extracts contain a heat-stable, nondialyzable activity that will inhibit the spontaneous assembly in vitro of partially purified brain tubulin. The sensitivity of this inhibitory activity to ribonucleases but not to a variety of other hydrolytic enzymes indicates that the inhibitor is an RNA. This conclusion is supported by the observation that purified RNAs from sea urchins, chinese hamster ovary cells, and brain all inhibit spontaneous microtubule assembly in vitro. The synthetic polynucleotides [poly(A), (C), (G), and (U)] are also inhibitory. This inhibition, however, appears to be nonspecific since the RNA base composition is unimportant and a variety of other nonnucleic acid polyanions also function as inhibitors. The treatment of assembly competent tubulin preparations with an insoluble RNA in the form of poly(A) covalently linked to agarose beads produces a "stripped" tubulin which will not assemble microtubules unless a heat-stable, trypsin-sensitive fraction eluted with increased ionic strength is mixed with the "stripped" tubulin. Similar results can be obtained with other cation exchangers, including phosphocellulose and carboxymethylcellulose. The heat-stable protein sequestered by poly(A)-agarose appears to be identical to the "tau" factor recently described by Kirschner and coworkers. Reconstitution experiments indicate that there is a stoichiometric requirement for these factors. These results suggest that spontaneous assembly of microtubules in nonneural cell extracts is blocked because the endogenous factors are complexed with RNA. This idea is supported by the observation that the ratio of tubulin to RNA is low in cultured cell extracts but very high in neural tissue extracts. 相似文献