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1.
A study was conducted into the course of meiosis in F 1 interspecific hybrids of Lycopersicum esculentum Mill (mutant line Mo 638) × Lycopersicum chinense Dul. and its parental forms. An F 1 interspecific hybrid was obtained through the embryo culture technique. A decrease in the chiasma frequency and an increase in the frequency of univalents and meiotic abnormalities compared to their parental forms were detected in hybrid plants. The number of univalents and the percentage of main impairments decreased, as the height of bud tier locations increased. A conclusion was made regarding the connection between the regularity of meiosis in the examined F 1 interspecific hybrids of Lycopersicon esculentum × Lycopersicon chilense, on the one hand, and the hybrid nature of genotypes and the influence of environmental factors, on the other hand. 相似文献
3.
Intergeneric hybrids between Brassica juncea (2n=36), B. carinata (2n=34) and Orychophragmus violaceus (2n=24) were produced when B. juncea and B. carinata cultivars were used as female parents. The hybrids between B. juncea and O. violaceus had an intermediate morphology except for petal colour and were partially fertile. The hybrids between B. carinata and O. violaceus had a matroclinous morphology and were nearly fertile. Cytological analysis of the hybrids and their progenies gave the following
results. (1) In the hybrids between B. juncea and O. violaceus, the somatic tissues of the roots, leaves and styles were mixoploid (2n=12–42), and cells with 24, 30 or 36 chromosomes were
the most frequent. Based on the recorded numbers and behaviour of the mitotic and meiotic chromosomes, complete and partial
separation of the parental genomes was proposed to have occurred during mitosis. This resulted in the occurrence of cells
with possibly complete and incomplete complements of the parental species and cells with parental complements and some additional
chromosomes from the other parent. (2) Pollen mother cells (PMCs) possibly with both parental chromosome complements, only
B. juncea chromosomes or a complete B. juncea complement with additional O. violaceus chromosomes were more competitive in entering meiosis. The majority of fertile gametes were deduced to have been produced
by PMCs with a B. juncea complement with or without additional O. violaceus chromosomes. (3) The progeny plants from selfed hybrids between B. juncea and O. violaceus were morphologically either of a B. juncea, hybrid or variable type. Cytologically they were grouped into six types according to the frequencies of cells with various
chromosome numbers. All of the plants except 2 which constituted two types, were mixoploids, composed of cells with various
chromosome numbers, mainly in a certain serial range. (4) The hybrid plants between B. carinata and O. violaceus were mixoploids with chromosome numbers in the range of 12–34, and cells with 2n=34 were the most frequent. The main categories
of PMCs with 17 bivalents at metaphase I and 17 : 17 segregations at anaphase I contributed to the high fertility of the hybrids
and the fact that their progeny after selfing were mainly plants with 2n=34. Somatic and meiotic separation of the parental
genomes was proposed to have occurred in the hybrids between B. carinata and O. violaceus. (5) Mitotic and meiotic elimination of what could be O. violaceus chromosomes might also have contributed to the observed mitotic and meiotic cell types in the two kinds of hybrids studied.
Finally, the possible mechanisms behind these cytological observations and their potential in the production of Brassica aneuploids were discussed.
Received: 4 February 1997/Accepted: 29 July 1997 相似文献
4.
Tordyliopsis brunonis (Apiaceae) is cytologically investigated here for the first time from India. The chromosome count of 2 n = 33, ascertained here, represents a new intraspecific triploid cytotype in the species, supplementing the earlier report of a diploid cytotype with 2 n = 22 from Nepal Himalayas. The diploid chromosome count ( n = 11) has also been found in some of the presently investigated individuals which showed perfectly normal meiosis with 100 % pollen fertility and normal seed set. However, the individuals with triploid chromosome count showed irregular meiotic behaviour and abnormal microsporogenesis resulting in high pollen sterility (56.26 %) and no seed set. The irregular meiotic behaviour in the triploid individuals is attributed to the occurrence of variable number of univalents (1–7) at diakinesis and metaphase-I. In the subsequent meiotic stages, these univalents lagged at anaphases and constituted micronuclei in sporads. The triploid plants were also observed for natural propagation and it was noticed that no seeds were set. These plants were noticed to propagate vegetatively by rootstocks. Chromosomal pairing in triploid cytotype is typical of an allopolyploid. Based on the characterization of chromosomal pairing during meiosis, we assumed that the triploid individuals are probably alloploid in nature. Hypotheses concerning the possible origin of allotriploid in T. brunonis are also discussed. 相似文献
5.
Responses of caryopsis germination, seedling emergence, and development of Agropyron cristatum (L.) Gaertn. (Gramineae) and Bromus inermis Leyss. (Gramineae), two dominant perennial grasses in the Otindag Sandland of China, to different sand water content (SWC; 1%, 2%, 3%, 4%, 6%, 8%, 12%, 16%, and 20%) were studied comparatively. The results showed that the germination responses of the two grasses to SWC were similar (i.e. caryopses could not germinate when the SWC was below 3%; at SWC ranging from 3% to 12%, the higher the SWC, the higher the germination percentage; and at a SWC of 12%-20%, germination reached similarly high percentages). At a sand burial depth of 0.5 cm, the threshold of SWC for seedling emergence was 6% forA. cristatum and 8% forB. inermis; at 12%-20% SWC, the seedling emergence of both species reached similarly high percentages. The seedling growth responses of these two species to SWC gradients were different. For A. cristatum, the biomass of seedlings increased with SWC from 6% to 12%, and decreased with SWC from 12% to 20%. For B. inermis, the biomass of seedlings always increased with SWC from 8% to 20%. The results also showed that the seedlings of both species allocated more biomass to the roots with decreases in SWC. The SWC changes from April to October in natural microhabitats of both species suggested that the SWC may play an important role in caryopsis germination, seedling emergence, and the growth characteristics of the two grasses. The responses of caryopsis germination, seedling emergence, and the growth characteristics of these two species to SWC may determine their distribution pattems in the Otindag Sandland. 相似文献
6.
There is strong evidence indicating that gene flow from transgenic B. napus into weedy wild relatives is inevitable following commercial release. Research should now focus on the transmission, stability,
and impact of transgene expression after the initial hybridization event. The present study investigated the transfer of a
phosphinothricin-tolerance transgene by inter-specific hybridization between B. rapa and two transgenic B. napus lines. The expression of the transgene was monitored in the F 1 hybrids and in subsequent backcross generations. The transgene was transmitted relatively easily into the F 1 hybrids and retained activity. Large differences in the transmission frequency of the transgene were noted between offspring
of the two transgenic lines during backcrossing. The most plausible explanation of these results is that the line showing
least transmission during backcrossing contains a transgene integrated into a C-genome chromosome. Approximately 10% of offspring
retained the tolerant trait in the BC 3 and BC 4 generations. The implications of these findings for the stable introgression of transgenes carried on one of the chromosomes
of the C-genome from B. napus and into B. rapa are briefly discussed.
Received: 5 November 1996 / Accepted: 21 February 1997 相似文献
7.
This paper analyzes the interaction between science, philosophy and politics (including ideology) in the early work of J. B. S. Haldane (from 1922 to 1937). This period is particularly important, not only because it is the period of Haldane's most significant biological work (both in biochemistry and genetics), but also because it is during this period that his philosophical and political views underwent their most significant transformation. His philosophical stance first changed from a radical organicism to a position far more compatible with mechanical materialism. The primary intellectual influence that was responsible for this shift was that of F. G. Hopkins. Later, Haldane came to accept Marxism and its official metaphysics, dialectical materialism, a move that let him accept the materialist conception of the world while still maintaining a resolute distance from mechanism. Throughout all these changes, what is most obvious is the influence of science on Haldane's philosophical views. An influence in the opposite direction is far less apparent.Parts of this paper are extracted from a longer work which concerns the interactions between philosophy and science throughout Haldane's scientific career (Sarkar forthcoming). The general conclusions reached here, from a consideration of Haldane's work only from 1922 to 1937 (see Section 6), remain the same for the rest of his life, as is detailed in the longer work. Thanks are due to R. S. Cohen, J. F. Crow, A. R. Fersht, J. Maynard Smith, R. C. Olby, D. Paul, M. Ruse, J. Stachel and S. Sturdy for helpful discussions and comments and criticism of the positions outlined in this paper. This is Contribution No. BTBG-92-4 from the Theoretical Biology Group, Boston University. 相似文献
8.
Species of Broussonetia have been essential in the development of papermaking technology. In Japan and Korea, a hybrid between B. monoica and B. papyrifera (= B. × kazinoki) known as kōzo and daknamu is still the major source of raw materials for making traditional paper washi and hanji, respectively. Despite their cultural and practical significance, however, the origin and taxonomy of kōzo and daknamu remain controversial. Additionally, the long-held generic concept of Broussonetia s.l., which included Sect. Allaeanthus and Sect. Broussonetia, was challenged as phylogenetic analyses showed Malaisia is sister to the latter section. To re-examine the taxonomic proposition that recognizes Allaeanthus, Broussonetia, and Malaisia (i.e., Broussonetia alliance), plastome and nuclear ribosomal DNA (nrDNA) sequences of six species of the alliance were assembled. Characterized by the canonical quadripartite structure, genome alignments and contents of the six plastomes (160,121–162,594 bp) are highly conserved, except for the pseudogenization and/or loss of the rpl22 gene. Relationships of the Broussonetia alliance are identical between plastome and nrDNA trees, supporting the maintenance of Malaisia and the resurrection of Allaeanthus. The phylogenomic relationships also indicate that the monoecy in B. monoica is a derived state, possibly resulting from hybridization between the dioecious B. kaempferi (♀) and B. papyrifera (♂). Based on the hypervariable ndhF-rpl32 intergenic spacer selected by sliding window analysis, phylogeographic analysis indicates that B. monoica is the sole maternal parent of B. × kazinoki and that daknamu carries multiple haplotypes, while only one haplotype was detected in kōzo. Because hybridizations between B. monoica and B. papyrifera are unidirectional and have occurred rarely in nature, our data suggest that daknamu might have originated via deliberate hybrid breeding selected for making hanji in Korea. On the contrary, kōzo appears to have a single origin and the possibility of a Korean origin cannot be ruled out. 相似文献
9.
Brassica carinata, an allotetraploid with B and C genomes, has a number of traits that would be valuable to introgress into B. napus. Interspecific hybrids were created between B. carinata (BBCC) and B. napus (AACC), using an advanced backcross approach to identify and introgress traits of agronomic interest from the B. carinata genome and to study the genetic changes that occur during the introgression process. We mapped the B and C genomes of B. carinata with SSR markers and observed their introgression into B. napus through a number of backcross generations, focusing on a BC(3) and BC(3)S(1) sibling family. There was close colinearity between the C genomes of B. carinata and B. napus and we provide evidence that B. carinata C chromosomes pair and recombine normally with those of B. napus, suggesting that similar to other Brassica allotetraploids no major chromosomal rearrangements have taken place since the formation of B. carinata. There was no evidence of introgression of the B chromosomes into the A or C chromosomes of B. napus; instead they were inherited as whole linkage groups with the occasional loss of terminal segments and several of the B-genome chromosomes were retained across generations. Several BC(3)S(1) families were analyzed using SSR markers, genomic in situ hybridization (GISH) assays, and chromosome counts to study the inheritance of the B-genome chromosome(s) and their association with morphological traits. Our work provides an analysis of the behavior of chromosomes in an interspecific cross and reinforces the challenges of introgressing novel traits into crop plants. 相似文献
10.
Summary The Giemsa C-banding technique was used to identify individual meiotic and somatic chromosomes in 21 monosomic lines of Avena byzantina C. Koch cv Kanota (genome designation AACCDD). The hexaploid complement is composed of three sets of seven chromosome pairs. The heterochromatin in the putative diploid progenitors is located at the telomeres (genome A), at the centromeric and interstitial regions (genome C), or more evenly spread throughout the set (genome D). Comparisons based on C-banding between A. byzantina and its diploid progenitor species allowed us to allocate individual chromosomes into specific genomes. The C-banding technique may be useful for interspecific chromosome pairing analyses. Nucleolar activity and competition were studied using a silver-staining procedure. Only three chromosome pairs showed nucleolar organizer regions, thus indicating that nucleolar competition occurs naturally in hexaploid oats. 相似文献
11.
The karyotypes in spermatogonial and leukocyte metaphases of the toads Bufo bufo, B. viridis and B. calamita (all 2n=22) were analysed and the DNA content of colchicine treated and Feulgen stained spermatogonial metaphase chromosomes measured microspectrophotometrically. The toad species possess similar karyotypes, but the chromosomes of B. bufo are somewhat longer than the chromosomes of B. viridis and B. calamita. All chromosomes of B. bufo contain significantly more than, but in no case twice as much DNA as their homologues in the other two species. Eight chromosomes of B. bufo contain 30–40%, three about 50% more DNA than their homologues in B. viridis. Exactly the same DNA-differences between both sets of chromosomes were found in B. bufo × B. viridis hybrids. Significant differences in the DNA amount of B. viridis and B. calamita exist only between the large chromosomes of these species. The ratio of the total DNA amount of the genomes in the three species is 1.49∶1.07∶1. These DNA-differences between the three toad species are confirmed by microspectrophotometric DNA measurements of their erythrocyte nuclei. It is supposed that these interspecific differences in DNA content of the toads are not a consequence of differential polyteny but are caused during the evolution process by local increase in DNA in all chromosomes of B. bufo and in the large chromosomes of B. viridis. 相似文献
12.
Bacterial cell division is orchestrated by the divisome, a protein complex centered on the tubulin homolog FtsZ. FtsZ polymerizes into a dynamic ring that defines the division site, recruits downstream proteins, and directs peptidoglycan synthesis to drive constriction. Recent studies have documented treadmilling of FtsZ polymer clusters both in cells and in vitro. Emerging evidence suggests that FtsZ dynamics are regulated largely by intrinsic properties of FtsZ itself and by the membrane anchoring protein FtsA. Although FtsZ dynamics are broadly required for Z-ring assembly, their role(s) during constriction may vary among bacterial species. These recent advances set the stage for future studies to investigate how FtsZ dynamics are physically and/or functionally coupled to peptidoglycan metabolic enzymes to direct efficient division. 相似文献
13.
Summary A thoroughly documented account of the ultrastructure of the meiotic spindle pole body (SPB) cycle in a rust ( Basidiomycota, Uredinales) is presented for the first time. The three-dimensional structure of the SPB and spindle during meiosis in the hollyhock rust fungus Puccinia malvacearum is analyzed from serial sections of preselected stages. This paper covers prophase I to prometaphase I. At late prophase I, the nucleolus disperses and does not reappear until the end of meiosis. The SPB at late prophase I consists of two, 4-layered discs, 0.8–1.0 m in diameter, connected by a middle piece (MP). The SPB is associated with a differentiated region of the nuclear envelope and nucleoplasm. At late diplotene to diakinesis, each disc generates a half spindle as it inserts into an otherwise intact nuclear envelope. The MP connecting the interdigitating half spindles elongates and eventually splits transversely during subsequent spindle elongation. Each half MP, which is attached to a SPB disc, becomes inserted in a sheath-like extension of the nuclear envelope. The intranuclear late prometaphase I spindle always becomes oriented perpendicularly to the longitudinal axis and sagittal plane of the metabasidium. There are 200–290 spindle microtubules (MTs) at each SPB at late prometaphase. The nonkinetochore MTs form a coherent central spindle around which the kinetochore MTs and bivalents are spread. A metaphase plate is absent. The results are compared with SPB behavior and spindle structure in early meiosis of other basidiomycetes and ascomycetes. 相似文献
14.
Summary The three-dimensional structure of the spindle pole body (SPB) and meiotic spindle during early metaphase I through telophase I in Puccinia malvacearum is analyzed ultrastructurally from serial sections. During early metaphase I the spindle rotates from the perpendicular to a position oblique to the longitudinal axis and parallel to the sagittal plane of the cell. Tubular cisternae are present within the spindle at this stage. The half middle piece (MP) subtends a collateral disc (co-disc) which is inserted eccentrically within each SPB. The SPB, co-disc and half MP at opposite poles are in mirror image. During the transition from early metaphase I to full metaphase I, the spindle orients parallel to the lateral wall of the promycelium and the half MPs are lost. The co-discs partially detach from each discoid SPB and maintain this relation until the end of interphase I. Co-discs become further differentiated as they attach to the subtending sheath-like extension of the nuclear envelope previously occupied by the half MPs. Microvesicles within the nucleoplasm are specific to mid metaphase I. A metaphase plate is absent. The 14 bivalents, which are directly connected to each polar SPB by 2 to 3 kinetochore MTs, are spread over nearly the entire length of the central spindle. The first anaphasic movement involves asynchronous shortening of the kinetochore MTs while the second consists of extensive pole-to-pole elongation. Astral MTs first appear at early metaphase I and become most numerous at anaphase I. An intact nuclear envelope constricts against the central spindle at either end of the interzonal region. Concurrently, centripetal growth of the nuclear envelope under each SPB results in their gradual externalization by the end of telophase I. The sibling nuclei are cut off by constriction of the nuclear envelope at either end of the interzonal region. These meiotic stages in P. malvacearum are compared with those in other basidiomycetes and ascomycetes. 相似文献
15.
This study presents the results of experiments concerning: (1) interspecific hybridization of Brassica oleracea × Brassica rapa via application of in vitro placental pollination and (2) embryological analysis of the process of resynthesis of Brassica napus. In order to overcome certain stigma/style barriers, B. rapa pollen was placed in vitro on an opened B. oleracea ovary (with style removed). Pollinated ovaries were cultured on Murashige and Skoog (MS) medium. After 24-d culture, the developing embryos were isolated from immature seeds and transferred onto MS medium supplemented with 0.47 μM kinetin, 0.49 μM 1-naphthaleneacetic acid, and 10% ( v/ v) coconut water. When the embryos had turned green, they were immediately placed onto MS medium with 100 μM kinetin. After development of the seedling, plantlets were transferred to soil. Chromosome doubling was achieved after another week. Cytometric analysis of nuclear DNA confirmed the hybrid nature of the plants. Resynthesis of B. napus can be performed through interspecific hybridization of B. oleracea × B. rapa followed by embryo rescue and genome doubling. 相似文献
16.
Allopolyploidy plays an important role in plant evolution and confers obvious advantages on crop growth and breeding compared to low ploidy levels. The present investigation was aimed at synthesising the first known chromosomally stable hexaploid Brassica with the genome constitution AABBCC. More than 2,000 putative hexaploid plants were obtained through large-scale hybridisation from various combinations of crosses between different cultivars of Brassica carinata (BBCC) and B. rapa (AA). The majority of plants after two generations of selfing within selected hexaploid plants (H 2) were aneuploid, and only 80 plants (4.6%) had the expected hexaploid chromosome number (2 n = 54). The hexaploid ratio increased to an average of 23.0 and 26.3% in the H 3 and H 4 generations, respectively, and was accompanied by an increase in pollen fertility. The appearance of aneuploid plants in each generation could be detected having various chromosomal abnormalities at meiosis. The frequency of hexaploid plants varied significantly among different cultivar combinations, from 0 to 56% in the H 4 generation, and it showed a positive correlation with pollen fertility. The frequency of SSR allelic fragments lost or novel alleles gained was significantly lower in H 4 than in H 2 and H 3, which reflects increasing genome stability in H 4. The A and C genomes were significantly less stable than the B genome, which may mainly result from frequent homoeologous pairing and rearrangements between the A and C genomes. Methods to establish a stable hexaploid Brassica crop by intercrossing these lines followed by intensive selection are also discussed. 相似文献
17.
1. Rates of entry and oxidation of a range of metabolites have been measured in tracheostomized sheep (diet, 800g. of lucerne chaff and 100g. of maize/day) by combining isotope-dilution techniques with the continuous measurement of total respiratory gas exchange, and 14CO 2 production during the intravenous or intraruminal infusion of 14C-labelled substrates. 2. Mean entry rates in fed and starved (24hr.) sheep respectively, expressed as mg./min./kg. body wt. 0·75, were: glucose, 5·0 (range 4·8–5·1, 2 observations) and 3·8 (3·2–4·2, 4); acetate, 10·8 (9·1–13·5, 4) and 5·8 (1); d(−)-β-hydroxybutyrate, 1·4 (1) and 1·5 (0·8–2·4, 4); palmitate, oleate and stearate (starved sheep only) 1·0 (0·6–1·9, 7), 0·9 (0·2–1·6, 10) and 0·9 (0·5–1·1, 11) respectively. 3. Production rates of propionate and butyrate in continuously feeding sheep were 6·4 (4·7–8·3, 4) and 4·3 (3·4–6·1, 4) mg./min./kg. 0·75 respectively, and in starved (24hr.) sheep were 2·5 (2·2–2·9, 2) and 1·0 (0·8–1·2, 2) mg./min./kg. 0·75 respectively. 4. Calculated terminal values for the specific radioactivity of respiratory 14CO 2 during measurements of entry rates and production rates were used to calculate the contributions of individual substrates to overall oxidative metabolism. Mean values for fed and starved sheep respectively were: glucose, 9·1 (8·6–9·6, 2) and 11·2 (5·9–15·1, 4)%; acetate, 31·6 (26·8–38·1, 4) and 22·1 (1)%; d(−)-β-hydroxybutyrate, 10·4 (1) and 4·8 (1·9–7·7, 4)%; propionate, 23·0 (13·8–29·9, 4) and 7·1 (6·8–7·4, 2)%; butyrate, 16·5 (13·7–20·5, 4) and 5·3 (5·2–5·3, 2)%; palmitate, oleate and stearate (starved sheep only), 4·7 (2·0–7·7, 7), 4·0 (1·2–6·6, 10) and 4·4 (3·8–5·8, 9)% respectively. The sum of these values for individual substrates in fed and starved sheep, excluding that of β-hydroxybutyrate and after correction of the glucose value for the known interrelations of this substrate with propionate, accounted for 76% and 58% respectively of total production of carbon dioxide. 5. Calculations based on the proportion of substrate entry directly oxidized indicated that the substrates studied accounted for 63% (fed sheep) and 43% (starved sheep) of total energy expenditure measured by oxygen uptake. The contribution of β-hydroxybutyrate was excluded, and corrections were made for glucose–propionate interrelations, and for the different rates of oxidation of the methyl and carboxyl fragments of acetate. 6. The present results have been combined with those obtained earlier in this Laboratory to examine the relationships between rates of substrate entry and oxidation, and concentrations of substrate in blood. Rates of entry of acetate, glucose, d(−)-β-hydroxybutyrate, palmitate and oleate (but not stearate) were well correlated with concentration in blood, and substrate contribution to production of carbon dioxide showed a similar correlation to blood concentration, except with glucose. 7. It was concluded that the general technique is of potential value in providing valid quantitative parameters of animal metabolism. 相似文献
18.
Chromosome pairing and chiasma frequency were studied in meiocytes at diakinesis of Lilium speciosum cv. Rosemede fixed up to 21 days after the start of either continuous or 3 day pulse colchicine treatment. The two treatments gave similar results. In pulse treated pollen mother cells (PMCs) the mean chiasma frequency per cell fell from 26.4 in controls to 8.5 after fourteen days while the mean number of univalents per cell increased from 0.05 to 17.58. There was a negative correlation between mean chiasma frequency per bivalent and per PMC in colchicine treated buds; univalents were preferentially induced in bivalents with one chiasma, and preferentially excluded in bivalents with 4 chiasmata. Some chiasmata were redistributed to surviving bivalents despite the concurrent reduction in chiasma frequency per meiocyte. — Colchicine sensitivity began in premeiotic interphase and extended to mid or late zygotene in PMCs; ongoing synapsis was unaffected. However, susceptibility to univalency was asynchronous between bivalents occurring at zygotene in short chromosomes but at late premeiotic interphase in the longest chromosomes. The number of chiasmata per bivalent could be altered by colchicine without inducing univalents, but the ultimate effect was to reduce the number of chiasmata per bivalent (or per chromosome arm) directly to zero. The major factors determining the order and extent of reduced pairing and chiasma number were total chromosome length and arm length. Pairing and chiasma formation in embryo sac mother cells were less sensitive to colchicine than in PMCs, but their behavior was otherwise similar. 相似文献
19.
During mitosis,the parent cell distributes its genetic materials equally into two daughter cells through chromosome segregation,a complex movements orchestrated by mitotic kinases and its effector proteins.Faithful chromosome segregation and cytokinesis ensure that each daughter cell receives a full copy of genetic materials of parent cell.Defects in these processes can lead to aneuploidy or polyploidy.Aurora/Ipllp family, a class of conserved serine/threonine kinases,plays key roles in chromosome segregation and cytokinesis.This article highlights the function and regulation of Aurora/Ipllp family in mitosis and provides potential links between aberrant regulation of Aurora/Ipllp kinases and pathogenesis of human cancer. 相似文献
20.
In the somatic cell, the mitotic spindle apparatus is centrosomal, and several isoforms of protein kinase C (PKC) have been
associated with the mitotic spindle, but their role in stabilizing the mitotic spindle is still unclear. Other protein kinases
such as, glycogen synthase kinase 3β (GSK3β) have also been shown to be associated with the mitotic spindle apparatus. In
this study, we show the enrichment of active (phosphorylated) PKCζ at the centrosomal region of the spindle apparatus in metaphase
stage of 3T3 cells. In order to understand whether the two kinases PKC and GSK3β are associated with the mitotic spindle,
first, the co-localization of phosphorylated PKC isoforms with GSK3β was studied at the poles in metaphase cells. Fluorescence
resonance energy transfer (FRET) analysis was used to demonstrate close molecular proximity of phospho-PKCζ with phospho(ser9)GSK3β.
Second, the involvement of inactive GSK3β in maintaining an intact mitotic spindle in 3T3 cells was shown. Third, this study
also showed that addition of a phospho-PKCζ specific inhibitor to cells can disrupt the mitotic spindle microtubules and some
of the proteins associated with it. The mitotic spindle at metaphase in mouse fibroblasts appears to be maintained by PKCζ
acting through GSK3β. Phospho-PKCζ is in close molecular proximity to GSK3β, whereas the other isoforms of PKC such as pPKCβII,
pPKCγ, pPKCμ, and pPKCθ are not close enough to have significant FRET readings. The close molecular proximity supports the
idea that GSK3β may be a substrate of PKCζ. 相似文献
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