DOCSLIB.ORG

Lymphocyte Subsets Cyclin D3-Cdk4

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Lymphocyte Subsets Cyclin D3-Cdk4 Cutting Edge: Differential Signaling Requirements for Activation of Assembled Cyclin D3-cdk4 Complexes in B-1 and B-2 Lymphocyte Subsets This information is current as of October 2, 2021. Debra A. Tanguay, Thomas P. Colarusso, Cheryl Doughty, Sandra Pavlovic-Ewers, Thomas L. Rothstein and Thomas C. Chiles J Immunol 2001; 166:4273-4277; ; doi: 10.4049/jimmunol.166.7.4273 Downloaded from http://www.jimmunol.org/content/166/7/4273 References This article cites 29 articles, 19 of which you can access for free at: http://www.jimmunol.org/content/166/7/4273.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on October 2, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. ● Cutting Edge: Differential Signaling Requirements for Activation of Assembled Cyclin D3-cdk4 Complexes in B-1 and B-2 Lymphocyte Subsets1 Debra A. Tanguay,2* Thomas P. Colarusso,3†§ Cheryl Doughty,* Sandra Pavlovic-Ewers,4* Thomas L. Rothstein,†‡§ and Thomas C. Chiles5* to the peritoneum, whereas B-2 cells predominate in spleen and Downloaded from B-1 lymphocytes represent a distinct B cell subset with unusual lymph nodes. B-1 cells contribute substantial proportions of non- mitogenic responses. PMA alone promotes proliferation in B-1 immune (resting) IgM and IgA that is repertoire restricted. cells, but not in splenic B-2 cells. Although cyclin D2-cyclin- Whether B-1 cells represent a developmental lineage distinct from dependent kinase 4 (cdk4) complexes mediate early retinoblas- B-2 cells or, alternatively, derive from a single B cell lineage in toma gene product (pRb) phosphorylation in B-1 cells, the which B-2 cells differentiate to B-1 cells in response to B cell Ag transient nature of their accumulation cannot account for the 6 receptor (BCR) -derived signals remains a matter of controversy http://www.jimmunol.org/ continued increase in pRb phosphorylation, which is maximal (4–6). Although B-1 cells resemble activated B-2 cells in terms of at 24 h. We show herein that PMA promotes the accumulation surface expression of IL-5R, CD44, and nuclear, activated STAT3 of functional cyclin D3-cdk4 complexes in B-1 cells following (7, 8), many additional molecular and transcriptional markers as- loss of cyclin D2. PMA also induces accumulation of cyclin sociated with B-2 cell activation are absent in B-1 cells (9, 10). D3-cdk4 complexes in B-2 cells; however, these complexes do B-1 cells differ significantly from B-2 cells in the signals re- not phosphorylate pRb. Thus, PMA is sufficient to induce syn- quired to induce proliferation. B-1 cells fail to enter S phase in thesis and assembly of cyclin D3-cdk4 complexes in B-1 and response to anti-Ig, whereas B-2 cells are mitogenically stimulated B-2 cells; however, PMA triggers cyclin D3-cdk4 activation by BCR cross-linking (11, 12). Treatment with phorbol ester alone only in B-1 cells. These results reveal a novel regulatory step is sufficient to stimulate B-1 cells to enter S phase, whereas B-2 by guest on October 2, 2021 that controls activation of cyclin D3-cdk4 complexes whose cells exit quiescence, but subsequently arrest in G1 phase of the function segregates differentially in B cell subsets. The Jour- cell cycle (progression to S phase requires a second signal pro- vided by calcium ionophore) (11, 12). The molecular basis under- nal of Immunology, 2001, 166: 4273–4277. lying the unique proliferative response of B-1 cells to phorbol esters is not completely understood. -1 cells constitute a unique subset of B lymphocytes, dis- It is generally considered that growth signals regulate mamma- tinguished from conventional B lymphocytes (B-2) by lian cell cycle entry by stimulating the accumulation of D-type B numerous phenotypic and functional characteristics (re- cyclins (cyclins D1, D2, and D3) that function to activate a subset viewed in Refs. 1–3). As an example, B-1 cells localize primarily of cyclin-dependent kinases (cdks) (reviewed in Ref. 13). The ret- inoblastoma gene product (pRb) is a target of cdks and acts to suppress G1-to-S phase progression (14–16). pRb is presently the *Department of Biology, Boston College, Chestnut Hill, MA 02467; and Departments most plausible candidate for regulating progression through the † ‡ § of Medicine and Microbiology, and The Evans Memorial Department of Clinical restriction (R) point (14, 16). A current model holds that pRb sup- Research, Boston University Medical Center, Boston, MA 02118 pression is alleviated through hyperphosphorylation that is medi- Received for publication December 8, 2000. Accepted for publication January 31, 2001. ated by both cyclin E and D-type cyclin kinase complexes (15, 16). The costs of publication of this article were defrayed in part by the payment of page The proper timing and extent of cdk activation is controlled by charges. This article must therefore be hereby marked advertisement in accordance dephosphorylation of inhibitory sites, phosphorylation of activat- with 18 U.S.C. Section 1734 solely to indicate this fact. ing sites, the action of two families of cdk inhibitors (Ink4 and 1 This work was supported by Grant MCB-9603784 awarded by the National Science Cip/Kip family), and by cyclin binding (17). For example, D-type Foundation (to T.C.C.) and U.S. Public Health Service Grant AI29690 awarded by the National Institutes of Health (to T.L.R.). cyclins and cyclin E function as positive regulatory subunits for cdk4/6 and cdk2, respectively (13). The requirement of mamma- 2 Current address: Genaissance Pharmaceuticals, Five Science Park, New Haven, CT 06511. lian cyclins D1 and D2 in G1 phase progression has been defini- 3 Current address: Genetics Computer Group, 575 Science Drive, Madison, WI tively established (18, 19). In keeping with this, distinct pheno- Ϫ Ϫ Ϫ Ϫ 53719. types have been reported in cyclin D1 / and cyclin D2 / mice 4 Current address: Fred Hutchinson Cancer Research Center, Seattle, WA 98109. 5 Address correspondence and reprint requests to Dr. Thomas C. Chiles, Department of Biology, Boston College, 414 Higgins Hall, Chestnut Hill, MA 02467. E-mail 6 Abbreviations used in this paper: BCR, B cell Ag receptor; pRb, retinoblastoma address: [email protected] gene product; cdk, cyclin-dependent kinase; R point, restriction point. Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 ● 4274 CUTTING EDGE (20, 21). Recent studies suggest that cyclin D3 may function to material was removed by centrifugation, and the supernatant was incubated limit the rate of G phase progression (18, 22). Interestingly, in with 1.5 ␮g nonimmune rabbit IgG, 1.5 ␮g of anti-cdk4, or 1.5 ␮gof 1 ␮ cyclin D2Ϫ/Ϫ mice, B-2 cells appear to remain responsive to mi- anti-cdk6 Abs. After 3 h, 50 l of a 1:1 slurry of protein G-agarose was added and incubated for 1 h. The immune complexes were recovered by togenic signals due to a compensatory induction of cyclin D3 lev- centrifugation and washed six times with Rb buffer and then three times in els (23). a buffer of 50 mM HEPES, pH 7.4, and 1 mM DTT. The immune Cell cycle progression to S phase in normal B-2 cells requires complexes were resuspended in 30 ␮l of Rb kinase buffer (50 mM HEPES, the accumulation of cyclin D2 and cdk4 and to a lesser extent cdk6 pH 7.5, 10 mM MgCl2, 5 mM MnCl2, 1 mM DTT, 2.5 mM EGTA, 10 mM ␤ ␮ ␥ 32 -glycerophosphate, 0.1 mM Na3VO4, and 10 Ci [ P]ATP at 6000 (24–26). B-2 cells from xid mice, which exhibit aborted activation Ci/mmol) in the presence of 1 ␮g of a truncated Rb protein substrate in response to BCR cross-linking, do not up-regulate cyclin D2 (p56Rb). After 15 min at 30°C, the reactions were terminated by the protein, suggesting that accumulation of cyclin D2 in B-2 cells addition of 2ϫ SDS sample buffer, and the reaction products were sepa- may be linked to passage through the R point (26). Interestingly, rated through a 10% polyacrylamide SDS gel. Phosphorylated Rb was Solvason et al. (27) recently demonstrated a requirement for cyclin detected by autoradiography of the dried gel. D2 expression in CD5 B cell development. We have previously Immunoblot analysis demonstrated that phorbol ester induces unusually early and tran- sient expression of cyclin D2 in B-1, but not in B-2 cells (28). As For the detection of cyclins D2 and D3, B lymphocytes were solubilized in 100 ␮l Triton X-100 buffer (50 mM HEPES, pH 7.4, 15 mM EGTA, 137 such, we proposed that the early induction of cyclin D2 may ac- ␤ mM NaCl, 15 mM MgCl2, 0.1% Triton X-100, 10 mM -glycerophos- ␮ count for the rapid entry of B-1 cells into the cell cycle following phate, 1 mM Na3VO4, 1 mM PMSF, and 1 g/ml aprotinin/leupeptin); for phorbol ester stimulation.
Load more

Recommended publications
  • Requirement for Cyclin D3 in Germinal Center Formation and Function
    Cell Research (2010) :1-16. © 2010 IBCB, SIBS, CAS All rights reserved 1001-0602/10 $ 32.00 npg ORIGINAL ARTICLE www.nature.com/cr Requirement for cyclin D3 in germinal center formation and function Jonathan U Peled1, J Jessica Yu1, Jeganathan Venkatesh2, Enguang Bi1, B Belinda Ding1, 5, Melissa Krupski-Downs1, Rita Shaknovich3, Piotr Sicinski4, Betty Diamond2, Matthew D Scharff1, B Hilda Ye1 1Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA; 2The Center for Autoimmune and Musculoskeletal Disease, The Feinstein Institute for Medical Research, Manhasset, NY 11030, USA; 3Depart- ments of Medicine and Pathology, Weill Cornell College of Medicine, New York, NY 10021, USA; 4Department of Cancer Biology, Dana-Farber Cancer Institute, and Department of Pathology, Harvard Medical School, Boston, MA 02115, USA Germinal centers (GC) of secondary lymphoid tissues are critical to mounting a high-affinity humoral immune response. B cells within the GC undergo rapid clonal expansion and selection while diversifying their antibody genes. Although it is generally believed that GC B cells employ a unique proliferative program to accommodate these pro- cesses, little is known about how the GC-associated cell cycle is orchestrated. The D-type cyclins constitute an im- portant component of the cell cycle engine that enables the cells to respond to physiological changes. Cell type- and developmental stage-specific roles of D-type cyclins have been described but the cyclin D requirement during GC reaction has not been addressed. In this study, we report that cyclin D3 is largely dispensable for proliferation and Ig class switching of in vitro activated B cells.
    [Show full text]
  • Altered Gene Expression Encoding Cytochines, Grow Factors and Cell Cycle Regulators in the Endometrium of Women with Chronic Endometritis
    diagnostics Article Altered Gene Expression Encoding Cytochines, Grow Factors and Cell Cycle Regulators in the Endometrium of Women with Chronic Endometritis Ettore Cicinelli 1, Amerigo Vitagliano 2,* , Vera Loizzi 1, Dominique De Ziegler 3, Margherita Fanelli 4 , Stefano Bettocchi 1, Claudia Nardelli 1, Giuseppe Trojano 1, Rossana Cicinelli 1, Crescenzio Francesco Minervini 5 , Daniela Leronni 6 and Luigi Viggiano 7 1 2nd Unit of Obstetrics and Gynecology, Department of Biomedical and Human Oncologic Science, Policlinico University of Bari, 70124 Bari, Italy; [email protected] (E.C.); [email protected] (V.L.); [email protected] (S.B.); [email protected] (C.N.); [email protected] (G.T.); [email protected] (R.C.) 2 Department of Women and Children’s Health, University of Padua, 35128 Padua, Italy 3 Department of Ob Gyn and Reproductive Medicine, Foch Hospital, 92150 Suresnes, France; [email protected] 4 Interdisciplinary Department of Medicine, University of Bari, 70124 Bari, Italy; [email protected] 5 Department of Emergency and Organ Transplantation (D.E.T.O.), Hematology Section, University of Bari, 70124 Bari, Italy; [email protected] 6 Neuroapoptosis Laboratory, Department of Neurological Surgery, University of Pittsburgh, Pittsburgh, PA 15213, USA; [email protected] 7 Department of Biology, University of Bari, 700124 Bari, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-333-1467105; Fax: +39-049-8211785 Citation: Cicinelli, E.; Vitagliano, A.; Loizzi, V.; De Ziegler, D.; Fanelli, M.; Abstract: To evaluate the expression of genes encoding cytokines, grow factors and cell cycle regula- Bettocchi, S.; Nardelli, C.; Trojano, G.; tors in the proliferative endometrium of women with chronic endometritis (CE) compared to controls.
    [Show full text]
  • Open Data for Differential Network Analysis in Glioma
    International Journal of Molecular Sciences Article Open Data for Differential Network Analysis in Glioma , Claire Jean-Quartier * y , Fleur Jeanquartier y and Andreas Holzinger Holzinger Group HCI-KDD, Institute for Medical Informatics, Statistics and Documentation, Medical University Graz, Auenbruggerplatz 2/V, 8036 Graz, Austria; [email protected] (F.J.); [email protected] (A.H.) * Correspondence: [email protected] These authors contributed equally to this work. y Received: 27 October 2019; Accepted: 3 January 2020; Published: 15 January 2020 Abstract: The complexity of cancer diseases demands bioinformatic techniques and translational research based on big data and personalized medicine. Open data enables researchers to accelerate cancer studies, save resources and foster collaboration. Several tools and programming approaches are available for analyzing data, including annotation, clustering, comparison and extrapolation, merging, enrichment, functional association and statistics. We exploit openly available data via cancer gene expression analysis, we apply refinement as well as enrichment analysis via gene ontology and conclude with graph-based visualization of involved protein interaction networks as a basis for signaling. The different databases allowed for the construction of huge networks or specified ones consisting of high-confidence interactions only. Several genes associated to glioma were isolated via a network analysis from top hub nodes as well as from an outlier analysis. The latter approach highlights a mitogen-activated protein kinase next to a member of histondeacetylases and a protein phosphatase as genes uncommonly associated with glioma. Cluster analysis from top hub nodes lists several identified glioma-associated gene products to function within protein complexes, including epidermal growth factors as well as cell cycle proteins or RAS proto-oncogenes.
    [Show full text]
  • Selective Usage of D-Type Cyclins in Lymphoid Malignancies
    Leukemia (1999) 13, 1335–1342 1999 Stockton Press All rights reserved 0887-6924/99 $15.00 http://www.stockton-press.co.uk/leu Selective usage of D-type cyclins in lymphoid malignancies R Suzuki1,2, H Kuroda1, H Komatsu1, Y Hosokawa1, Y Kagami2, M Ogura2, S Nakamura3, Y Kodera4, Y Morishima2, R Ueda5 and M Seto1 1Laboratory of Chemotherapy, 2Department of Hematology and Chemotherapy, and 3Department of Pathology and Clinical Laboratories, Aichi Cancer Center; 4Department of Internal Medicine, Japanese Red Cross Nagoya First Hospital; and 5Second Department of Internal Medicine, Nagoya City University School of Medicine, Nagoya, Japan Three D-type cyclins, cyclin D1, D2 and D3, belong to the G1 some 14,11–13 in much the same way as the c-myc gene is cyclin, which regulates the G1/S transition of the cell cycle, and affected by t(8;14) translocations in Burkitt’s lymphoma14 and feature highly homologous amino acid sequences. The cyclin the BCL-2 gene by t(14;18) translocations in follicular lym- D1 gene was found to be transcriptionally activated in B-lymph- 15 oid malignancies with t(11;14), but available information is lim- phomas. The breakpoints occur at variable distances from ited regarding expression of cyclin D2 and D3 in hematopoietic the cyclin D1 gene, typically up to 120 kb, but the net effect malignancies. We examined the expressions of three D-type appears to be the transcriptional activation of cyclin cyclins to investigate how these homologous genes are differ- D1.12,13,16–18 Several groups including ours have reported entially used.
    [Show full text]
  • Cytometry of Cyclin Proteins
    Reprinted with permission of Cytometry Part A, John Wiley and Sons, Inc. Cytometry of Cyclin Proteins Zbigniew Darzynkiewicz, Jianping Gong, Gloria Juan, Barbara Ardelt, and Frank Traganos The Cancer Research Institute, New York Medical College, Valhalla, New York Received for publication January 22, 1996; accepted March 11, 1996 Cyclins are key components of the cell cycle pro- gests that the partner kinase CDK4 (which upon ac- gression machinery. They activate their partner cy- tivation by D-type cyclins phosphorylates pRB com- clin-dependent kinases (CDKs) and possibly target mitting the cell to enter S) is perpetually active them to respective substrate proteins within the throughout the cell cycle in these tumor lines. Ex- cell. CDK-mediated phosphorylation of specsc sets pression of cyclin D also may serve to discriminate of proteins drives the cell through particular phases Go vs. GI cells and, as an activation marker, to iden- or checkpoints of the cell cycle. During unper- tify the mitogenically stimulated cells entering the turbed growth of normal cells, the timing of expres- cell cycle. Differences in cyclin expression make it sion of several cyclins is discontinuous, occurring possible to discrirmna* te between cells having the at discrete and well-defined periods of the cell cy- same DNA content but residing at different phases cle. Immunocytochemical detection of cyclins in such as in G2vs. M or G,/M of a lower DNA ploidy vs. relation to cell cycle position (DNA content) by GI cells of a higher ploidy. The expression of cyclins multiparameter flow cytometry has provided a new D, E, A and B1 provides new cell cycle landmarks approach to cell cycle studies.
    [Show full text]
  • D3-Deficient Mice Is Compensated by Cyclin D2 in Cyclin of Normal B-1A
    Disruption of Cyclin D3 Blocks Proliferation of Normal B-1a Cells, but Loss of Cyclin D3 Is Compensated by Cyclin D2 in Cyclin D3-Deficient Mice This information is current as of September 28, 2021. Jennifer M. Mataraza, Joseph R. Tumang, Maria R. Gumina, Sean M. Gurdak, Thomas L. Rothstein and Thomas C. Chiles J Immunol 2006; 177:787-795; ; doi: 10.4049/jimmunol.177.2.787 Downloaded from http://www.jimmunol.org/content/177/2/787 References This article cites 51 articles, 21 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/177/2/787.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 28, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Disruption of Cyclin D3 Blocks Proliferation of Normal B-1a Cells, but Loss of Cyclin D3 Is Compensated by Cyclin D2 in Cyclin D3-Deficient Mice1 Jennifer M.
    [Show full text]
  • Cyclin D2 and Cyclin D3 Play Opposite Roles in Mouse Skin Carcinogenesis
    Oncogene (2007) 26, 1723–1730 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE Cyclin D2 and cyclin D3 play opposite roles in mouse skin carcinogenesis P Rojas1,4, MB Cadenas2,4, P-C Lin3, F Benavides1, CJ Conti1 and ML Rodriguez-Puebla2 1Department of Carcinogenesis, Science Park Research Division, MD Anderson Cancer Center, Smithville, TX, USA; 2Center for Comparative Medicine and Translational Research, Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA and 3Department of Molecular and Cellular Oncology, MD Anderson Cancer Center, Houston, TX, USA D-type cyclins are components ofthe cell-cycle engine a family of key cell-cycle regulators, as they are the that link cell signaling pathways and passage throughout regulatory subunits of the CDK4 and CDK6, which G1 phase. We previously described the effects of over- phosphorylate pRb family of proteins that are critical expression cyclin D1, D2 or D3 in mouse epidermis and substrates for cell-cycle progression (Matsushine et al., tumor development. We now asked whether cyclin D2 1994; Meyerson and Harlow, 1994; Sherr and Roberts, and/or cyclin D3 play a relevant role in ras-dependent 1999; Farkas et al., 2002; Leng et al., 2002). Initially, tumorigenesis. Here, we described the effect of cyclin D3 several reports assigned redundant roles to the three and cyclin D2 overexpression in mouse skin tumor members of D-type cyclins, but in the last few years, it development. Notably, overexpression ofcyclin D3 results has become evident that each member is differentially in reduced tumor development and malignant progression expressed in various tissues playing specific roles (Sherr, to squamous cell carcinomas (SCC).
    [Show full text]
  • Full Text (PDF)
    654 Vol. 1, 654–664, July 2003 Molecular Cancer Research Expression of Cyclin-Dependent Kinase 6, but not Cyclin-Dependent Kinase 4, Alters Morphology of Cultured Mouse Astrocytes Karen K. Ericson, David Krull, Peter Slomiany, and Martha J. Grossel Department of Biology, Connecticut College, New London, CT Abstract mechanism involving the accumulation of the p53 and p130 Disruption of the pRb pathway is a common mechanism growth-suppressing proteins (6), and activation of cdk6 in tumor formation. The D-cyclin-associated kinases, precedes cdk4 activation in T cells (7, 8). Also, differences in cyclin-dependent kinase (cdk) 4 and cdk6, are important subcellular localization of cdk4 and cdk6 and in the timing of regulators of the G1-S phase transition and are elevated nuclear localization have been noted in several cell types by in several types of cancers, including gliomas. To several researchers (9–12). Data from tumor studies also investigate potential functional differences in these suggest differences in these kinases. Cdk4, and not cdk6, is kinases, mouse astrocytes were taken from chimeric specifically targeted in melanoma (13, 14) while cdk6 activity mice and propagated in tissue culture. These multipolar has been found to be elevated in squamous cell carcinomas (15, tissue-culture astrocytes were infected with viruses 16) and neuroblastomas (17) without alteration of cdk4 activity. expressing either cdk4 or cdk6. Interestingly, expression Cumulatively, these data suggest that cdk4 and cdk6 have of cdk6 resulted in a distinct and rapid morphology unique functions that may be cell-type specific, temporally change from multipolar to bipolar. This change was not regulated, or developmentally distinct.
    [Show full text]
  • New Functional Activities for the P21 Family of CDK Inhibitors
    Downloaded from genesdev.cshlp.org on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press New functional activities for the p21 family of CDK inhibitors Joshua LaBaer/'* Michelle D. Garrett,^ Lauren F. Stevenson/ Joyce M. Slingerland,^ Charanjit Sandhu,^ Hubert S. Chou/ All Fattaey/ and Ed Harlow^ ^Massachusetts General Hospital Cancer Center, Charlestown, Massachusetts 02129; ^Onyx Pharmaceuticals, Richmond, California 94806 USA; '^Division of Cancer Biology Research, Sunnybrook Health Science Center, S-207, Toronto, Ontario M4N 3M5 Canada The association of cdk4 with D-type cyclins to form functional kinase complexes is comparatively inefficient. This has led to the suggestion that assembly might be a regulated step. In this report we demonstrate that the CDK inhibitors pZl'^^'', p27^^^, and p57^^^^ all promote the association of cdk4 with the D-type cyclins. This effect is specific and does not occur with other cdk inhibitors or cdk-binding proteins. Both in vivo and in vitro, the abundance of assembled cdk4/cyclin D complex increases directly with increasing inhibitor levels. The promotion of assembly is not attributable to a simple cell cycle block and requires the function of both the cdk and cyclin-binding domains. Kinetic studies demonstrate that p21 and p27 lead to a 35- and 80-fold increase in K^, respectively, mostly because of a decrease in X^ff. At low concentrations, p21 promotes the assembly of active kinase complexes, whereas at higher concentrations, it inhibits activity. Moreover, immunodepletion experiments demonstrate that most of the active cdk4-associated kinase activity also associates with p21. To confirm these results in a natural setting, we examine the assembly of endogenous complexes in mammary epithelial cells after release from a GQ arrest.
    [Show full text]
  • Sonic Hedgehog-Dependent Activation of Gli2 Is Essential for Embryonic Hair Follicle Development
    Downloaded from genesdev.cshlp.org on September 29, 2021 - Published by Cold Spring Harbor Laboratory Press Sonic hedgehog-dependent activation of Gli2 is essential for embryonic hair follicle development Pleasantine Mill,1 Rong Mo,1 Hong Fu,1 Marina Grachtchouk,2 Peter C.W. Kim,3 Andrzej A. Dlugosz,2 and Chi-chung Hui1,4 1Program in Developmental Biology, The Hospital for Sick Children, and Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario M5G 1X8, Canada; 2University of Michigan Comprehensive Cancer Center and Department of Dermatology, Ann Arbor, Michigan 48109, USA; 3Program in Infection, Immunity and Repair, The Hospital for Sick Children, and Department of Surgery, University of Toronto, Toronto, Ontario M5G 1X8, Canada Sonic hedgehog (Shh) signaling plays a critical role in hair follicle development and skin cancer, but how it controls these processes remains unclear. Of the three Gli transcription factors involved in transducing Shh signals in vertebrates, we demonstrate here that Gli2 is the key mediator of Shh responses in skin. Similar to Shh−/− mice, Gli2−/− mutants exhibit an arrest in hair follicle development with reduced cell proliferation and Shh-responsive gene expression, but grossly normal epidermal differentiation. By transgenic rescue experiments, we show that epidermal Gli2 function alone is sufficient to restore hair follicle development in Gli2−/− skin. Furthermore, only a constitutively active form of Gli2, but not wild-type Gli2, can activate Shh-responsive gene expression and promote cell proliferation in Shh−/− skin. These observations indicate that Shh-dependent Gli2 activator function in the epidermis is essential for hair follicle development. Our data also reveal that Gli2 mediates the mitogenic effects of Shh by transcriptional activation of cyclin D1 and cyclin D2 in the developing hair follicles.
    [Show full text]
  • Differentiation Regulated Differently During Human B Cell Cyclin
    The Expression of p18INK4 and p27kip1 Cyclin-Dependent Kinase Inhibitors Is Regulated Differently During Human B Cell Differentiation1 Nicolas Schrantz,2 Georges Edouard Beney,2 Marie The´re`se Auffredou, Marie Franc¸oise Bourgeade, Ge´rald Leca, and Aime´Vazquez3 Cell cycle progression is under the control of cyclin-dependent kinases (cdks), the activity of which is dependent on the expression of specific cdk inhibitors. In this paper we report that the two cdk inhibitors, p27Kip1 and p18INK4c, are differently expressed and control different steps of human B lymphocyte activation. Resting B cells contain large amounts of p27Kip1 and no p18INK4c.In vitro stimulation by Staphylococcus aureus Cowan 1 strain or CD40 ligand associated with IL-10 and IL-2 induces a rapid decrease in p27Kip1 expression combined with cell cycle entry and progression. In contrast, in vitro Ig production correlates with specific INK4c expression of p18 and early G1 arrest. This G1 arrest is associated with inhibition of cyclin D3/cdk6-mediated retinoblastoma protein phosphorylation by p18INK4c. A similar contrasting pattern of p18INK4c and p27Kip1 expression is observed both in B cells activated in vivo and in various leukemic cells. Expression of p18INK4c was also detected in various Ig-secreting cell lines in which INK4c Kip1 both maximum Ig secretion and specific p18 expression were observed during the G1 phase. Our study shows that p27 and p18INK4c have different roles in B cell activation; p27Kip1 is involved in the control of cell cycle entry, and p18INK4c is involved in the subsequent early G1 arrest necessary for terminal B lymphocyte differentiation.
    [Show full text]
  • The Role of APC (Anaphase-Promoting Comlpex) in G2/M After DNA Damage Jinho Lee
    The role of APC (Anaphase-Promoting Comlpex) in G2/M after DNA damage Jinho Lee To cite this version: Jinho Lee. The role of APC (Anaphase-Promoting Comlpex) in G2/M after DNA damage. Cellular Biology. Université Joseph-Fourier - Grenoble I, 2007. English. tel-00247413 HAL Id: tel-00247413 https://tel.archives-ouvertes.fr/tel-00247413 Submitted on 8 Aug 2009 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. UNIVERSITE JOSEPH FOURIER-GRENOBLE 1 Sciences, Technologie & Médicine THESE pour obtenir le grade de DOCTEUR DE L’UNIVERSITE JOSEPH FOURIER SPECIALITE : BIOLOGIE CELLULAIRE Présenté et soutenue publiquement par Jinho LEE Le rôle du APC (Anaphase-Promoting Complex) au cours de la phase G2/M après dommage de l’ADN The role of APC (Anaphase-Promoting Comlpex) in G2/M after DNA damage INSTITUT DE BIOLOGIE STRUCTURALE Jean-Pierre Ebel (Université Joseph Fourier) Laboratoire des Proteines du Cytosquelette Grenoble, France SIDNEY KIMMEL CANCER CENTER Cancer Cell Biology Program San Diego, USA Directeur de thèse : Dr. Rati Fotedar Pr. Michel Robert-Nicoud Président Dr. Paolo Sassone-Corsi Rapporteur Dr. Rakesh Kumar Rapporteur Dr. Robert Margolis Examinateur Dr. Rati Fotedar Directrice de thèse et examinatrice Le 29 Octobre 2007 au Sidney Kimmel Cancer Center Remerciements D’abord, je voudrais exprimer ma reconnaissance à tous ceux qui m’ont permis de réaliser ma thèse et à toutes les assistances financières pour mon étude en France.
    [Show full text]