title: Reshaping of the endoplasmic reticulum limits the rate for nuclear envelope formation.
abstract: During mitosis in metazoans, segregated chromosomes become enclosed by the nuclear envelope (NE), a double membrane that is continuous with the endoplasmic reticulum (ER). Recent in vitro data suggest that NE formation occurs by chromatin-mediated reorganization of the tubular ER; however, the basic principles of such a membrane-reshaping process remain uncharacterized. Here, we present a quantitative analysis of nuclear membrane assembly in mammalian cells using time-lapse microscopy. From the initial recruitment of ER tubules to chromatin, the formation of a membrane-enclosed, transport-competent nucleus occurs within approximately 12 min. Overexpression of the ER tubule-forming proteins reticulon 3, reticulon 4, and DP1 inhibits NE formation and nuclear expansion, whereas their knockdown accelerates nuclear assembly. This suggests that the transition from membrane tubules to sheets is rate-limiting for nuclear assembly. Our results provide evidence that ER-shaping proteins are directly involved in the reconstruction of the nuclear compartment and that morphological restructuring of the ER is the principal mechanism of NE formation in vivo.
authors.affiliation: Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Anderson
authors.forename: Martin W
authors.initials: MW
authors.lastname: Hetzer
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D000070798:Nogo Proteins; D009685:Nuclear Envelope / Q000378:metabolism* / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000032:analysis; D050680:Transcription Factor DP1 / Q000235:genetics / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D002843:Chromatin; D008565:Membrane Proteins; D009185:Myelin Proteins; D009419:Nerve Tissue Proteins; D000070798:Nogo Proteins; C476198:RTN3 protein, human; C000603199:RTN4 protein, human; D011993:Recombinant Fusion Proteins; D050680:Transcription Factor DP1; D049452:Green Fluorescent Proteins
references.citation: Cell. 2006 Feb 10;124(3):573-86
references.pmid: 16469703
references.citation: J Cell Biol. 1997 Sep 22;138(6):1193-206
references.pmid: 9298976
references.citation: Genome Biol. 2007;8(12):234
references.pmid: 18177508
references.citation: Curr Opin Cell Biol. 2003 Feb;15(1):88-95
references.pmid: 12517709
references.citation: EMBO J. 2001 Mar 15;20(6):1320-30
references.pmid: 11250898
references.citation: Trends Cell Biol. 2006 Feb;16(2):67-9
references.pmid: 16406617
references.citation: J Cell Biol. 1991 Feb;112(4):545-56
references.pmid: 1993730
references.citation: Nat Cell Biol. 2007 Jul;9(7):822-31
references.pmid: 17558394
references.citation: EMBO J. 1996 Dec 16;15(24):7108-19
references.pmid: 9003786
references.citation: Methods. 1999 Nov;19(3):362-72
references.pmid: 10579931
references.citation: Science. 2007 Nov 30;318(5855):1408-12
references.pmid: 18048680
references.citation: J Cell Biol. 1992 Jan;116(2):281-94
references.pmid: 1730756
references.citation: J Cell Biol. 1977 Aug;74(2):492-500
references.pmid: 885912
references.citation: J Cell Biochem. 2005 Jun 1;95(3):454-65
references.pmid: 15832341
references.citation: J Cell Biol. 2006 May 22;173(4):469-76
references.pmid: 16717124
references.citation: J Cell Biol. 2006 Jan 2;172(1):41-53
references.pmid: 16380439
references.citation: Nat Cell Biol. 2007 Oct;9(10):1160-6
references.pmid: 17828249
references.citation: Trends Biochem Sci. 2004 Apr;29(4):175-82
references.pmid: 15082311
references.citation: J Cell Biol. 2007 Jul 2;178(1):43-56
references.pmid: 17591921
references.citation: Nat Cell Biol. 2001 Dec;3(12):1086-91
references.pmid: 11781570
references.citation: Cell Mol Life Sci. 2006 Feb;63(3):316-32
references.pmid: 16389459
references.citation: Curr Opin Cell Biol. 2008 Aug;20(4):386-92
references.pmid: 18495454
references.citation: Nat Cell Biol. 2004 Nov;6(11):1114-21
references.pmid: 15502822
references.citation: J Cell Biol. 2007 Dec 3;179(5):895-909
references.pmid: 18056408
references.citation: Annu Rev Cell Dev Biol. 2007;23:321-44
references.pmid: 17506702
references.citation: Science. 2006 Apr 21;312(5772):440-3
references.pmid: 16627745
references.citation: Subcell Biochem. 2000;34:273-302
references.pmid: 10808336
references.citation: Nat Biotechnol. 2004 Dec;22(12):1567-72
references.pmid: 15558047
references.citation: Cell. 2006 Nov 3;127(3):635-48
references.pmid: 17081983
references.citation: J Cell Biol. 2001 Jul 9;154(1):71-84
references.pmid: 11448991
references.citation: Science. 2008 Feb 29;319(5867):1247-50
references.pmid: 18309084
references.citation: Traffic. 2000 Sep;1(9):689-94
references.pmid: 11208156
references.citation: Nat Rev Mol Cell Biol. 2004 Jan;5(1):65-9
references.pmid: 14663490
references.citation: Cell. 2006 Aug 11;126(3):435-9
references.pmid: 16901774
references.citation: J Biol Chem. 2008 Jul 4;283(27):18892-904
references.pmid: 18442980