We examined mitotic chromosomes of three newly found members of the Drosophila obscura species group,D.luguensis,D.dianensis and D.limingi from Yunnan and compared their karyotypes to those of respective related speci...We examined mitotic chromosomes of three newly found members of the Drosophila obscura species group,D.luguensis,D.dianensis and D.limingi from Yunnan and compared their karyotypes to those of respective related species.D.luguensis possessed a diploid number of 12 chromosomes,comprising 3 pairs of metacentrics (V-shaped),2 pairs of acrocentrics (rod-like) and 1 pair of micro-chromosomes (dot-like).Both the X and Y were metacentrics.D.dianensis and D.limingi possessed a diploid number of 10 chromosomes,comprising 1 pair of large V-shaped,1 pair of moderate-sized V-shaped,2 pairs of submetacentric (J-shaped) chromosomes,and 1 pairs of dot-like chromosomes,with their X chromosomes were J-shaped and Y chromosomes were short rod-like ones.Based upon the results of chromosomal comparisons,geographic information of the sinobscura subgroup,as well inter-specific phylogenetic relationship deduced elsewhere,D.luguensis was considered to retain an ancestral form of the karyotype within this subgroup.The present observations have also implied that D.sinobscura (2n=12:2V,1J,2R,1D) might have derived from a sub-lineage of a pre-“sinobscura-hubeiensis” lineage by pericentric inversion of moderate-sized metacentric autosomes and D.hubeiensis (2n=10:4V,1D) from another sub-lineage of the same lineage by centric fusion of 2 pairs of acrocentric autosomes.Similar chromosomal changes of centric fusion,centric fission and/or pericentric inversion might have occurred during the course of species divergences between D.dianensis and D.subsilvestris (2n=12:3V,2R,1D),an European member closely related to D.dianensis,and between D.limingi and D.tsukubaensis (2n=12:3V,2R,1D),an East Asian member near to D.limingi.展开更多
To date,limited research has been carried out in developing methods and materials that offer three-dimensional(3-D) representation of the digestive tract.In the field of capsule endoscopy(CE),hardware approaches have ...To date,limited research has been carried out in developing methods and materials that offer three-dimensional(3-D) representation of the digestive tract.In the field of capsule endoscopy(CE),hardware approaches have been developed that provide real time both 3-D information and texture using an infrared projector and a complementary metal oxide semiconductor camera.The major drawbacks of this system are its size,power consumption and packaging issues.A software approach to approximate a 3-D representation of digestive tract surface utilising current CE technology has been proposed.The algorithm utilizes the Shape from Shading technique and seem to provide promising results for polypoid structures and angioectasias.Further clinical evaluation is currently under way.展开更多
Mitosin/CENP-F is a human nuclear protein transiently associated with the outer kinetochore plate in M phase and is involved in M phase progression. LEK1 and CMF1, which are its murine and chicken orthologs, however, ...Mitosin/CENP-F is a human nuclear protein transiently associated with the outer kinetochore plate in M phase and is involved in M phase progression. LEK1 and CMF1, which are its murine and chicken orthologs, however, are implicated in muscle differentiation and reportedly not distributed at kinetochores.We therefore conducted several assays to clarify this issue. The typical centromere staining patterns were observed in mitotic cells from both human primary culture and murine, canine, and mink cell lines. A C-terminal portion of LEK1 also conferred centromere localization. Our analysis further suggests conserved kinetochore localization of mammalian mitosin orthologs. Moreover, mitosin was associated preferentially with kinetochores of unaligned chromosomes. It was also constantly transported from kinetochores to spindle poles by cytoplasmic dynein. These properties resemble those of other kinetochore proteins important for the spindle checkpoint, thus implying a role of mitosin in this checkpoint. Therefore, mitosin family may serve as multifunctional proteins involved in both mitosis and differentiation.展开更多
For proper chromosome segregation, all kinetochores must achieve bipolar microtubule （MT） attachment and subsequently align at the spindle equator before anaphase onset. The MT minus end-directed motor dynein/dynact...For proper chromosome segregation, all kinetochores must achieve bipolar microtubule （MT） attachment and subsequently align at the spindle equator before anaphase onset. The MT minus end-directed motor dynein/dynactin binds kinetoehores in prometaphase and has long been implicated in chromosome congression. Unfortunately, inactivation of dynein usually disturbs spindle organization, thus hampering evaluation of its kinetochore roles. Here we specifically eliminated kinetochore dynein/dynactin by RNAi-mediated depletion of ZW10, a protein essential for kinetochore localization of the motor. Time-lapse microscopy indicated markedly-reduced congression efficiency, though congressing chromosomes displayed similar velocities as in control cells. Moreover, cells frequently failed to achieve full chromosome alignment, despite their normal spindles. Confocal microcopy revealed that the misaligned kinetochores were monooriented or unattached and mostly lying outside the spindle, suggesting a difficulty to capture MTs from the opposite pole. Kinetoehores on monoastral spindles were dispersed farther away from the pole and exhibited only mild oscillation. Furthermore, inactivating dynein by other means generated similar phenotypes. Therefore, kinetochore dynein produces on monooriented kinetochores a poleward pulling force, which may contribute to efficient bipolar attachment by facilitating their proper microtubule captures to promote congression as well as full chromosome alignment.展开更多
The aim of the present investigation was to study the major chromosomal aberrations （CA） like deletion, translocation, inversion and mosaic in prostate cancer patients of Tamilnadu, Southern India. Totally 45 blood ...The aim of the present investigation was to study the major chromosomal aberrations （CA） like deletion, translocation, inversion and mosaic in prostate cancer patients of Tamilnadu, Southern India. Totally 45 blood samples were collected from various hospitals in Tamilnadu, Southern India. Equal numbers of normal healthy subjects were chosen after signing a consent form. Volunteers provided blood samples （5 ml） to establish leukocyte cultures. Cytogenetic studies were performed by using Giemsaanding technique and finally the results were ensured by spectral karyotyping （SKY） technique. In the present investigation, major CA like deletion, translocation, inversion and mosaic were identifed in experimental subjects. Results showed frequent CA in chromosomes 1, 3, 5, 6, 7, 9, 13, 16, 18 and X. In comparison with experimental subjects, the control subjects exhibited very low levels of major CA （P〈0.05）. In the present study, the high frequency of centromeric rearrangements indicates a potential role for mitotic irregularities associated with the centromere in prostate cancer tumorigenesis. Identification of chromosome alterations may be helpful in understanding the molecular basis of the disease in better manner.展开更多
Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Our recent study shows that NEK2A interacts with MAD 1 at the kinetochore and possibly functions as a ...Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Our recent study shows that NEK2A interacts with MAD 1 at the kinetochore and possibly functions as a novel integrator of spindle checkpoint signaling. However, it is unclear how NEK2A regulates kinetochore-microtubule attachment in mitosis. Here we show that NEK2A phosphorylates human Sgo 1 and such phosphorylation is essential for faithful chromosome congression in mitosis. NEK2A binds directly to HsSgol in vitro and co-distributes with HsSgol to the kinetochore of mitotic cells. Our in vitro phosphorylation experiment demonstrated that HsSgo 1 is a substrate of NEK2A and the phosphorylation sites were mapped to Ser^14 and Ser^507 as judged by the incorporation of 32^P. Although such phosphorylation is not required for assembly of HsSgo 1 to the kinetochore, expression of non-phosphorylatable mutant HsSgo 1 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic and monotelic attachments. These findings reveal a key role for the NEK2A-mediated phosphorylation ofHsSgo 1 in orchestrating dynamic kinetochore-microtubule interaction. We propose that NEK2A-mediated phosphorylation of human Sgo 1 provides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores.展开更多
Cyclin B1 is a key regulatory protein controlling cell cycle progression in vertebrates. Cyclin B1 binds CDK1, a cy-clin-dependent kinase catalytic subunit, forming a complex that orchestrates mitosis through phosphor...Cyclin B1 is a key regulatory protein controlling cell cycle progression in vertebrates. Cyclin B1 binds CDK1, a cy-clin-dependent kinase catalytic subunit, forming a complex that orchestrates mitosis through phosphorylation of key proteins. Cyclin B1 regulates both the activation of CDK1 and its subcellular localization, which may be critical for substrate selection. Here, we demonstrate that cyclin B1 is concentrated on the outer plate of the kinetochore during prometaphase. This localization requires the cyclin box region of the protein. Cyclin B1 is displaced from individual kinetochores to the spindle poles by microtubule attachment to the kinetochores, and this displacement is dependent on the dynein/dynactin complex. Depletion of cyclin B1 by vector-based siRNA causes inefficient attachment between kinetochores and microtubules, and chromosome alignment defects, and delays the onset of anaphase. We conclude that cyclin B1 accumulates at kinetochores during prometaphase, where it contributes to the correct attachment of mi- crotubules to kinetochores and efficient alignment of the chromosomes, most likely through localized phosphorylation of specific substrates by cyclin B1-CDK1. Cyclin B1 is then transported from each kinetochore as microtubule attachment is completed, and this relocalization may redirect the activity of cyclin B1-CDK1 and contribute to inactivation of the spindle assembly checkpoint.展开更多
Recently the antichromosome antisera from several scleroderma patients have been found to recognize the pellicle of metaphase and anaphase chromosomes. In order to identify the pellicle components, we used these antic...Recently the antichromosome antisera from several scleroderma patients have been found to recognize the pellicle of metaphase and anaphase chromosomes. In order to identify the pellicle components, we used these antichromosome antisera to screen a human embryonic cDNA library. The sequences of the positive clones are identical to the cDNA gene sequence of CENP-C (centromere protein C), a human centromere autoantigen. This result suggusts that CENP-C is a component of the pellicle of human metaphase and anaphase chromosomes.展开更多
文摘We examined mitotic chromosomes of three newly found members of the Drosophila obscura species group,D.luguensis,D.dianensis and D.limingi from Yunnan and compared their karyotypes to those of respective related species.D.luguensis possessed a diploid number of 12 chromosomes,comprising 3 pairs of metacentrics (V-shaped),2 pairs of acrocentrics (rod-like) and 1 pair of micro-chromosomes (dot-like).Both the X and Y were metacentrics.D.dianensis and D.limingi possessed a diploid number of 10 chromosomes,comprising 1 pair of large V-shaped,1 pair of moderate-sized V-shaped,2 pairs of submetacentric (J-shaped) chromosomes,and 1 pairs of dot-like chromosomes,with their X chromosomes were J-shaped and Y chromosomes were short rod-like ones.Based upon the results of chromosomal comparisons,geographic information of the sinobscura subgroup,as well inter-specific phylogenetic relationship deduced elsewhere,D.luguensis was considered to retain an ancestral form of the karyotype within this subgroup.The present observations have also implied that D.sinobscura (2n=12:2V,1J,2R,1D) might have derived from a sub-lineage of a pre-“sinobscura-hubeiensis” lineage by pericentric inversion of moderate-sized metacentric autosomes and D.hubeiensis (2n=10:4V,1D) from another sub-lineage of the same lineage by centric fusion of 2 pairs of acrocentric autosomes.Similar chromosomal changes of centric fusion,centric fission and/or pericentric inversion might have occurred during the course of species divergences between D.dianensis and D.subsilvestris (2n=12:3V,2R,1D),an European member closely related to D.dianensis,and between D.limingi and D.tsukubaensis (2n=12:3V,2R,1D),an East Asian member near to D.limingi.
文摘To date,limited research has been carried out in developing methods and materials that offer three-dimensional(3-D) representation of the digestive tract.In the field of capsule endoscopy(CE),hardware approaches have been developed that provide real time both 3-D information and texture using an infrared projector and a complementary metal oxide semiconductor camera.The major drawbacks of this system are its size,power consumption and packaging issues.A software approach to approximate a 3-D representation of digestive tract surface utilising current CE technology has been proposed.The algorithm utilizes the Shape from Shading technique and seem to provide promising results for polypoid structures and angioectasias.Further clinical evaluation is currently under way.
基金supported by grants 97JC14006 from Shanghai Committee of Science and Technology(No.30025021,39970160,and 39500030)from the Natural Science Foundation of ChinaKSCX2-2-02 from Chinese Academy of Sciences.
文摘Mitosin/CENP-F is a human nuclear protein transiently associated with the outer kinetochore plate in M phase and is involved in M phase progression. LEK1 and CMF1, which are its murine and chicken orthologs, however, are implicated in muscle differentiation and reportedly not distributed at kinetochores.We therefore conducted several assays to clarify this issue. The typical centromere staining patterns were observed in mitotic cells from both human primary culture and murine, canine, and mink cell lines. A C-terminal portion of LEK1 also conferred centromere localization. Our analysis further suggests conserved kinetochore localization of mammalian mitosin orthologs. Moreover, mitosin was associated preferentially with kinetochores of unaligned chromosomes. It was also constantly transported from kinetochores to spindle poles by cytoplasmic dynein. These properties resemble those of other kinetochore proteins important for the spindle checkpoint, thus implying a role of mitosin in this checkpoint. Therefore, mitosin family may serve as multifunctional proteins involved in both mitosis and differentiation.
基金Acknowledgments The authors thank Qiongping Huang, Lirong Liu, and Wei Bian for technical assistance. We are grateful to Drs G Chan （Cross Cancer Institute, University of Alberta, Edmonton Alberta, Canada） for antibodies to human ZW 10 and Rod, KH Choo （Murdoch Children＇s Research Institute, Royal Children＇s Hospital, Melbourne, Australia） for anti-CREST serum, and E Fuchs （Rockefeller University, USA） for mRFP cDNA. This work was supported by the National Science Foundation of China （30330330, 30421005, and 30623003）, Ministry of Science and Technology of China （2005CB522703 and 2007CB914501）, and the Shanghai Municipal Council for Science and Technology （S048014317, 06DZ22032, and 058014578）.
文摘For proper chromosome segregation, all kinetochores must achieve bipolar microtubule （MT） attachment and subsequently align at the spindle equator before anaphase onset. The MT minus end-directed motor dynein/dynactin binds kinetoehores in prometaphase and has long been implicated in chromosome congression. Unfortunately, inactivation of dynein usually disturbs spindle organization, thus hampering evaluation of its kinetochore roles. Here we specifically eliminated kinetochore dynein/dynactin by RNAi-mediated depletion of ZW10, a protein essential for kinetochore localization of the motor. Time-lapse microscopy indicated markedly-reduced congression efficiency, though congressing chromosomes displayed similar velocities as in control cells. Moreover, cells frequently failed to achieve full chromosome alignment, despite their normal spindles. Confocal microcopy revealed that the misaligned kinetochores were monooriented or unattached and mostly lying outside the spindle, suggesting a difficulty to capture MTs from the opposite pole. Kinetoehores on monoastral spindles were dispersed farther away from the pole and exhibited only mild oscillation. Furthermore, inactivating dynein by other means generated similar phenotypes. Therefore, kinetochore dynein produces on monooriented kinetochores a poleward pulling force, which may contribute to efficient bipolar attachment by facilitating their proper microtubule captures to promote congression as well as full chromosome alignment.
文摘The aim of the present investigation was to study the major chromosomal aberrations （CA） like deletion, translocation, inversion and mosaic in prostate cancer patients of Tamilnadu, Southern India. Totally 45 blood samples were collected from various hospitals in Tamilnadu, Southern India. Equal numbers of normal healthy subjects were chosen after signing a consent form. Volunteers provided blood samples （5 ml） to establish leukocyte cultures. Cytogenetic studies were performed by using Giemsaanding technique and finally the results were ensured by spectral karyotyping （SKY） technique. In the present investigation, major CA like deletion, translocation, inversion and mosaic were identifed in experimental subjects. Results showed frequent CA in chromosomes 1, 3, 5, 6, 7, 9, 13, 16, 18 and X. In comparison with experimental subjects, the control subjects exhibited very low levels of major CA （P〈0.05）. In the present study, the high frequency of centromeric rearrangements indicates a potential role for mitotic irregularities associated with the centromere in prostate cancer tumorigenesis. Identification of chromosome alterations may be helpful in understanding the molecular basis of the disease in better manner.
基金We thank members of our group for insightful discussion during the course of this study.This work was supported by grants from Chinese Academy of Science(KSCX1-YW-R65,KSCX2-YW-H10)National Basic Research Program of China(2002CB713700)+4 种基金Hi-Tech Research and Development Program of China(2001AA215331)Chinese Minister of Education(20020358051 to XY,PCSIRT0413 to XD)National Natural Science Foundation of China(39925018,30270293 to XY,30500183 to XD,30600222 to JY)National Institutes of Health(USA)(DK56292,CA92080)to XY(a Georgia Cancer Coalition Eminent Scholar)JY was supported by China Postdoctor(2005037560).
文摘Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Our recent study shows that NEK2A interacts with MAD 1 at the kinetochore and possibly functions as a novel integrator of spindle checkpoint signaling. However, it is unclear how NEK2A regulates kinetochore-microtubule attachment in mitosis. Here we show that NEK2A phosphorylates human Sgo 1 and such phosphorylation is essential for faithful chromosome congression in mitosis. NEK2A binds directly to HsSgol in vitro and co-distributes with HsSgol to the kinetochore of mitotic cells. Our in vitro phosphorylation experiment demonstrated that HsSgo 1 is a substrate of NEK2A and the phosphorylation sites were mapped to Ser^14 and Ser^507 as judged by the incorporation of 32^P. Although such phosphorylation is not required for assembly of HsSgo 1 to the kinetochore, expression of non-phosphorylatable mutant HsSgo 1 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic and monotelic attachments. These findings reveal a key role for the NEK2A-mediated phosphorylation ofHsSgo 1 in orchestrating dynamic kinetochore-microtubule interaction. We propose that NEK2A-mediated phosphorylation of human Sgo 1 provides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores.
文摘Cyclin B1 is a key regulatory protein controlling cell cycle progression in vertebrates. Cyclin B1 binds CDK1, a cy-clin-dependent kinase catalytic subunit, forming a complex that orchestrates mitosis through phosphorylation of key proteins. Cyclin B1 regulates both the activation of CDK1 and its subcellular localization, which may be critical for substrate selection. Here, we demonstrate that cyclin B1 is concentrated on the outer plate of the kinetochore during prometaphase. This localization requires the cyclin box region of the protein. Cyclin B1 is displaced from individual kinetochores to the spindle poles by microtubule attachment to the kinetochores, and this displacement is dependent on the dynein/dynactin complex. Depletion of cyclin B1 by vector-based siRNA causes inefficient attachment between kinetochores and microtubules, and chromosome alignment defects, and delays the onset of anaphase. We conclude that cyclin B1 accumulates at kinetochores during prometaphase, where it contributes to the correct attachment of mi- crotubules to kinetochores and efficient alignment of the chromosomes, most likely through localized phosphorylation of specific substrates by cyclin B1-CDK1. Cyclin B1 is then transported from each kinetochore as microtubule attachment is completed, and this relocalization may redirect the activity of cyclin B1-CDK1 and contribute to inactivation of the spindle assembly checkpoint.
文摘Recently the antichromosome antisera from several scleroderma patients have been found to recognize the pellicle of metaphase and anaphase chromosomes. In order to identify the pellicle components, we used these antichromosome antisera to screen a human embryonic cDNA library. The sequences of the positive clones are identical to the cDNA gene sequence of CENP-C (centromere protein C), a human centromere autoantigen. This result suggusts that CENP-C is a component of the pellicle of human metaphase and anaphase chromosomes.
