title: The activity of a wall-bound cellulase is required for and is coupled to cell cycle progression in the dinoflagellate Crypthecodinium cohnii.
abstract: Cellulose synthesis, but not its degradation, is generally thought to be required for plant cell growth. In this work, we cloned a dinoflagellate cellulase gene, dCel1, whose activities increased significantly in G(2)/M phase, in agreement with the significant drop of cellulose content reported previously. Cellulase inhibitors not only caused a delay in cell cycle progression at both the G(1) and G(2)/M phases in the dinoflagellate Crypthecodinium cohnii, but also induced a higher level of dCel1p expression. Immunostaining results revealed that dCel1p was mainly localized at the cell wall. Accordingly, the possible role of cellulase activity in cell cycle progression was tested by treating synchronized cells with exogenous dCelp and purified antibody, in experiments analogous to overexpression and knockdown analyses, respectively. Cell cycle advancement was observed in cells treated with exogenous dCel1p, whereas the addition of purified antibody resulted in a cell cycle delay. Furthermore, delaying the G(2)/M phase independently with antimicrotubule inhibitors caused an abrupt and reversible drop in cellulase protein level. Our results provide a conceptual framework for the coordination of cell wall degradation and reconstruction with cell cycle progression in organisms with cell walls. Since cellulase activity has a direct bearing on the cell size, the coupling between cellulase expression and cell cycle progression can also be considered as a feedback mechanism that regulates cell size.
authors.affiliation: Department of Biology, Hong Kong University of Science and Technology, Clearwater Bay, Kowloon, Hong Kong SAR, People's Republic of China.
authors.forename: Alvin C M
authors.initials: AC
authors.lastname: Kwok
authors.forename: Joseph T Y
authors.initials: JT
authors.lastname: Wong
mesh_terms: D000595:Amino Acid Sequence; D002453:Cell Cycle; D002473:Cell Wall / Q000737:chemistry; D002482:Cellulose / Q000096:biosynthesis; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000235:genetics / Q000378:metabolism*; D004141:Dinoflagellida / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016053:RNA, Protozoan / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D016053:RNA, Protozoan; D002482:Cellulose; D043366:Cellulose 1,4-beta-Cellobiosidase
references.citation: Plant Physiol. 2001 Oct;127(2):529-42
references.pmid: 11598227
references.citation: Microbiol Rev. 1995 Sep;59(3):345-86
references.pmid: 7565410
references.citation: Science. 1989 Nov 3;246(4930):629-34
references.pmid: 2683079
references.citation: FEMS Microbiol Rev. 1994 Jan;13(1):25-58
references.pmid: 8117466
references.citation: Plant Cell Physiol. 2002 Dec;43(12):1399-406
references.pmid: 12514237
references.citation: Nature. 2009 Jun 11;459(7248):852-6
references.pmid: 19474792
references.citation: Plant Physiol. 2001 May;126(1):278-88
references.pmid: 11351091
references.citation: Anal Biochem. 1983 Jun;131(2):333-6
references.pmid: 6193735
references.citation: Plant Physiol. 1995 Jan;107(1):87-100
references.pmid: 11536663
references.citation: Plant Physiol. 1996 Jul;111(3):813-20
references.pmid: 8754682
references.citation: Science. 2002 Jan 4;295(5552):147-50
references.pmid: 11778054
references.citation: Exp Cell Res. 2000 Jan 10;254(1):120-9
references.pmid: 10623472
references.citation: Anal Biochem. 1987 Jul;164(1):72-7
references.pmid: 2445222
references.citation: Appl Environ Microbiol. 1997 Apr;63(4):1298-306
references.pmid: 9097427
references.citation: Protein Expr Purif. 2008 Mar;58(1):162-7
references.pmid: 17942322
references.citation: J Biol Chem. 1995 Oct 27;270(43):26012-9
references.pmid: 7592793
references.citation: Bioinformatics. 2007 Nov 1;23(21):2947-8
references.pmid: 17846036
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Eukaryot Cell. 2002 Oct;1(5):774-86
references.pmid: 12455695
references.citation: Crit Rev Biotechnol. 1989;9(2):61-103
references.pmid: 2509081
references.citation: Biochem Biophys Res Commun. 1995 Apr 26;209(3):1046-52
references.pmid: 7733957
references.citation: Gene. 2007 Jun 15;395(1-2):29-39
references.pmid: 17408885
references.citation: Plant Physiol. 2003 Apr;131(4):1681-91
references.pmid: 12692327
references.citation: Eur J Biochem. 2003 Aug;270(16):3455-60
references.pmid: 12899703
references.citation: Plant J. 2000 Oct;24(2):147-58
references.pmid: 11069690
references.citation: Plant Physiol. 2001 Sep;127(1):324-33
references.pmid: 11553760
references.citation: Plant J. 1993 Jan;3(1):1-30
references.pmid: 8401598
references.citation: PLoS Biol. 2005 Jul;3(7):e225
references.pmid: 15966770
references.citation: Science. 1996 Dec 6;274(5293):1664-72
references.pmid: 8939848
references.citation: Mol Biol Evol. 1987 Jul;4(4):406-25
references.pmid: 3447015
references.citation: Appl Microbiol Biotechnol. 1995 Dec;44(3-4):399-404
references.pmid: 8597541
references.citation: Sci China C Life Sci. 2004 Feb;47(1):18-24
references.pmid: 15382672
references.citation: Microbiology (Reading). 2001 Apr;147(Pt 4):781-794
references.pmid: 11283274
references.citation: Arch Biochem Biophys. 1995 Sep 10;322(1):119-26
references.pmid: 7574665
references.citation: EMBO J. 1998 Oct 1;17(19):5563-76
references.pmid: 9755157
references.citation: J Biol Chem. 2001 Jun 8;276(23):19679-82
references.pmid: 11309404
references.citation: J Biol Chem. 1984 Aug 25;259(16):10455-9
references.pmid: 6432782
references.citation: Anal Biochem. 1969 Dec;32(3):420-4
references.pmid: 5361396
references.citation: Nat Cell Biol. 2004 Sep;6(9):861-71
references.pmid: 15311283
references.citation: Plant Cell. 1995 Jul;7(7):987-1000
references.pmid: 7640530
references.citation: Plant Physiol. 1994 Feb;104(2):557-62
references.pmid: 8159787
references.citation: Annu Rev Plant Physiol Plant Mol Biol. 1999;50:391-417
references.pmid: 11541953
references.citation: Eur J Biochem. 1989 Oct 1;184(3):553-9
references.pmid: 2806239
references.citation: Plant Physiol. 1998 Oct;118(2):333-9
references.pmid: 9765519
references.citation: J Cell Biol. 1995 May;129(3):739-49
references.pmid: 7730408
references.citation: Gene. 2003 Dec 4;321:73-81
references.pmid: 14636994
references.citation: Plant Cell. 2005 Dec;17(12):3378-89
references.pmid: 16284310
references.citation: Plant J. 2003 Feb;33(4):677-90
references.pmid: 12609041
references.citation: Science. 2004 Jul 16;305(5682):354-60
references.pmid: 15256663
references.citation: Biotechniques. 2006 Jul;41(1):35-6, 38, 40 passim
references.pmid: 16869511
references.citation: Curr Top Dev Biol. 1999;46:39-61
references.pmid: 10417876
references.citation: Biochim Biophys Acta. 1969 Nov 4;191(2):342-53
references.pmid: 5354265
references.citation: Plant Physiol. 1988 Dec;88(4):1257-62
references.pmid: 16666452
references.citation: Appl Environ Microbiol. 1990 Oct;56(10):3117-24
references.pmid: 2126700
references.citation: Mol Biol Evol. 2007 Aug;24(8):1596-9
references.pmid: 17488738
references.citation: J Bacteriol. 1983 Nov;156(2):828-36
references.pmid: 6195146
references.citation: Microbiology (Reading). 2004 Jul;150(Pt 7):2029-2035
references.pmid: 15256547
references.citation: Plant J. 2003 Mar;33(6):1099-106
references.pmid: 12631333
references.citation: Microbiol Mol Biol Rev. 2005 Jun;69(2):262-91
references.pmid: 15944456
references.citation: Curr Protoc Immunol. 2001 May;Chapter 8:8.10.1-8.10.21
references.pmid: 18432850
references.citation: Science. 2004 Dec 24;306(5705):2191; author reply 2191
references.pmid: 15618503
references.citation: Biotechnol Bioeng. 1990 Jul;36(3):275-87
references.pmid: 18595079
references.citation: Biotechnol Bioeng. 1992 Sep;40(6):663-71
references.pmid: 18601165
references.citation: Arch Biochem Biophys. 1999 Apr 1;364(1):61-6
references.pmid: 10087165
references.citation: Plant J. 2003 Mar;33(6):1087-97
references.pmid: 12631332
references.citation: Curr Opin Microbiol. 2004 Dec;7(6):617-23
references.pmid: 15556034
references.citation: Plant Cell. 2000 Jul;12(7):1137-52
references.pmid: 10899980
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11724-9
references.pmid: 12172008
references.citation: Proc Natl Acad Sci U S A. 1977 Aug;74(8):3466-70
references.pmid: 269406
references.citation: Nat Cell Biol. 2004 Sep;6(9):801-3
references.pmid: 15340443
references.citation: J Cell Biol. 1998 Sep 21;142(6):1519-32
references.pmid: 9744881
references.citation: Nucleic Acids Res. 1997 Dec 15;25(24):4876-82
references.pmid: 9396791
references.citation: Int Rev Cytol. 1983;82:151-80
references.pmid: 6684652
references.citation: J Mol Biol. 1990 Oct 5;215(3):403-10
references.pmid: 2231712
references.citation: J Biol Chem. 1991 Oct 15;266(29):19758-67
references.pmid: 1918080
references.citation: Appl Microbiol Biotechnol. 2007 Oct;76(5):1069-78
references.pmid: 17641888
references.citation: Nature. 2009 Jun 11;459(7248):857-60
references.pmid: 19474789