Cell cycle-dependent SUMO-1 conjugation to nuclear mitotic apparatus protein (NuMA)

Jae Sung Seo; Ha Na Kim; Sun-Jick Kim; Jiyoung Bang; Eun-A Kim; Ki Sa Sung; Hyun-Joo Yoon; Hae Yong Yoo; Cheol Yong Choi

doi:10.1016/j.bbrc.2013.11.107

Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport.

The Journal of Cell Biology ◽

10.1083/jcb.115.1.1 ◽

1991 ◽

Vol 115 (1) ◽

pp. 1-17 ◽

Cited By ~ 486

Author(s):

J Pines ◽

T Hunter

Keyword(s):

Cell Cycle ◽

Human Fibroblasts ◽

Nuclear Lamina ◽

Cyclin A ◽

Cyclin B1 ◽

Cell Fractionation ◽

Mitotic Apparatus ◽

Epithelial Tumor ◽

Mitotic Cyclin ◽

Cycle Dependent

We have used immunofluorescence staining to study the subcellular distribution of cyclin A and B1 during the somatic cell cycle. In both primary human fibroblasts and in epithelial tumor cells, we find that cyclin A is predominantly nuclear from S phase onwards. Cyclin A may associated with condensing chromosomes in prophase, but is not associated with condensed chromosomes in metaphase. By contrast, cyclin B1 accumulates in the cytoplasm of interphase cells and only enters the nucleus at the beginning of mitosis, before nuclear lamina breakdown. In mitotic cells, cyclin B1 associates with condensed chromosomes in prophase and metaphase, and with the mitotic apparatus. Cyclin A is degraded during metaphase and cyclin B1 is precipitously destroyed at the metaphase----anaphase transition. Cell fractionation and immunoprecipitation studies showed that both cyclin A and cyclin B1 are associated with PSTAIRE-containing proteins. The nuclear, but not the cytoplasmic form, of cyclin A is associated with a 33-kD PSTAIRE-containing protein. Cyclin B1 is associated with p34cdc2 in the cytoplasm. Thus we propose that the different localization of cyclin A and cyclin B1 in the cell cycle could be the means by which the two types of mitotic cyclin confer substrate specificity upon their associated PSTAIRE-containing protein kinase subunit.

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Cell cycle-dependent phosphorylation of the 77 kDa echinoderm microtubule-associated protein (EMAP) in vivo and association with the p34cdc2 kinase

Journal of Cell Science ◽

10.1242/jcs.109.12.2885 ◽

1996 ◽

Vol 109 (12) ◽

pp. 2885-2893 ◽

Cited By ~ 2

Author(s):

E. Brisch ◽

M.A. Daggett ◽

K.A. Suprenant

Keyword(s):

Cell Cycle ◽

Sea Urchin ◽

Time Course ◽

Mitotic Apparatus ◽

Sea Urchin Eggs ◽

Spindle Poles ◽

A Cell ◽

Cycle Dependent ◽

Phosphopeptide Mapping

The most abundant microtubule-associated protein in sea urchin eggs and embryos is the 77 kDa echinoderm microtubule-associated protein (EMAP). EMAP localizes to the mitotic spindle as well as the interphase microtubule array and is a likely target for a cell cycle-activated kinase. To determine if EMAP is phosphorylated in vivo, sea urchin eggs and embryos were metabolically labeled with 32PO4 and a monospecific antiserum was used to immunoprecipitate EMAP from 32P-labeled eggs and embryos. In this study, we demonstrate that the 77 kDa EMAP is phosphorylated in vivo by two distinct mechanisms. In the unfertilized egg, EMAP is constitutively phosphorylated on at least five serine residues. During the first cleavage division following fertilization, EMAP is phosphorylated with a cell cycle-dependent time course. As the embryo enters mitosis, EMAP phosphorylation increases, and as the embryo exits mitosis, phosphorylation decreases. During mitosis, EMAP is phosphorylated on 10 serine residues and two-dimensional phosphopeptide mapping reveals a mitosis-specific site of phosphorylation. At all stages of the cell cycle, a 33 kDa polypeptide copurifies with the 77 kDa EMAP, regardless of phosphorylation state. Antibodies against the cdc2 kinase were used to demonstrate that the 33 kDa polypeptide is the p34cdc2 kinase. The p34cdc2 kinase copurifies with the mitotic apparatus and immunostaining indicates that the p34cdc2 kinase is concentrated at the spindle poles. Models for the interaction of the p34cdc2 kinase and the 77 kDa EMAP are presented.

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Cell Cycle-Dependent Localization of Voltage-Dependent Calcium Channels and the Mitotic Apparatus in a Neuroendocrine Cell Line(AtT-20)

International Journal of Cell Biology ◽

10.1155/2009/487959 ◽

2009 ◽

Vol 2009 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

Karen J. Loechner ◽

Wendy C. Salmon ◽

Jie Fu ◽

Shipra Patel ◽

James T. McLaughlin

Keyword(s):

Cell Cycle ◽

Calcium Channels ◽

Intracellular Calcium ◽

Cell Line ◽

Time Lapse ◽

Mitotic Apparatus ◽

Calcium Dependent ◽

Voltage Dependent ◽

Specific Localization ◽

Cycle Dependent

Changes in intracellular calcium are necessary for the successful progression of mitosis in many cells. Both elevation and reduction in intracellular calcium can disrupt mitosis by mechanisms that remain ill defined. In this study we explore the role of transmembrane voltage-gated calcium channels (CaV channels) as regulators of mitosis in the mouse corticotroph cell line (AtT-20). We report that the nifedipine-sensitive isoform CaV1.2 is localized to the “poleward side” of kinetechores during metaphase and at the midbody during cytokinesis. A second nifedipine-sensitive isoform, CaV1.3, is present at the mid-spindle zone in telophase, but is also seen at the midbody. Nifedipine reduces the rate of cell proliferation, and, utilizing time-lapse microscopy, we show that this is due to a block at the prometaphase stage of the cell cycle. Using Fluo-4 we detect calcium fluxes at sites corresponding to the mid-spindle zone and the midbody region. Another calcium dye, Fura PE3/AM, causes an inhibition of mitosis prior to anaphase that we attribute to a chelation of intracellular calcium. Our results demonstrate a novel, isoform-specific localization of CaV1 channels during cell division and suggest a possible role for these channels in the calcium-dependent events underlying mitotic progression in pituitary corticotrophs.

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Distribution of a matrix component of the midbody during the cell cycle in Chinese hamster ovary cells

The Journal of Cell Biology ◽

10.1083/jcb.106.2.431 ◽

1988 ◽

Vol 106 (2) ◽

pp. 431-439 ◽

Cited By ~ 87

Author(s):

C Sellitto ◽

R Kuriyama

Keyword(s):

Cell Cycle ◽

Chinese Hamster Ovary ◽

Chinese Hamster Ovary Cells ◽

Chinese Hamster ◽

Immunoblot Analysis ◽

Equatorial Region ◽

Dense Matrix ◽

Mitotic Apparatus ◽

Ovary Cells ◽

Cycle Dependent

Monoclonal antibodies were raised against isolated spindles of CHO (Chinese hamster ovary) cells to probe for molecular components specific to the mitotic apparatus. One of the antibodies, CHO1, recognized an antigen localized to the midbody during mitosis. Immunofluorescence staining of metaphase cells showed that although the total spindle area was labeled faintly, the antigen corresponding to CHO1 was preferentially localized in the equatorial region of the spindle. With the progression of mitosis, the antigen was further organized into discrete short lines along the spindle axis, and eventually condensed into a bright fluorescent dot at the midzone of the intercellular bridge between two daughter cells. Parallel immunostaining of tubulin showed that the CHO1-stained area corresponded to the dark region where microtubules are entrapped by the amorphous dense matrix components and possibly blocked from binding to tubulin antibody. Immunoblot analysis indicated that CHO1 recognized two polypeptides of mol wt 95,000 and 105,000. The immunoreaction was always stronger in preparations of isolated midbodies than in mitotic spindle fractions. The protein doublet was retained in the particulate matrix fraction after Sarkosyl extraction (Mullins, J. M., and J. R. McIntosh. 1982. J. Cell Biol. 94:654-661), suggesting that CHO1 antigen is indeed a component of the dense matrix. In addition to the equatorial region of spindles and midbodies, CHO1 also stained interphase centrosomes, and nuclei in a speckled pattern that was cell cycle-dependent. Thus, the midbody appears to share either common molecular component(s) or a similar epitope with interphase centrosomes and nuclei.

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