Cell Stem Cell
The Cell Cycle and Myc Intersect with Mechanismsthat Regulate Pluripotency and Reprogramming
Amar M. Singh
and Stephen Dalton
Department of Biochemistry and Molecular Biology, Paul D. Coverdell Center for Biomedical and Health Sciences,The University of Georgia, 500 D.W. Brooks Drive, Athens, GA 30602, USA *Correspondence:email@example.comDOI 10.1016/j.stem.2009.07.003
Pluripotentstemcellshavelong-termproliferativecapacityandanunusualmodeofcell-cycleregulationandcan divide independently of extrinsic mitogenic signals. The last few years has seen evidence emerge thatlinks cell-cycle regulation to the maintenance and establishment of pluripotency. Myc transcription factorsappear to be central to this regulation. This review addresses these links and discusses how cell-cyclecontrols and Myc impact on the maintenance and establishment of pluripotency.
ConnectionsbetweentheMycfamilyoftranscriptionfactorsandcell-cycle control have been of constant interest to biologists forover25years.Thisrelationshipisparticularlyevidentinthestudyof tumor biology, where deregulated Myc activity is known toplay a central role in aberrant cell proliferation and immortaliza-tion. More recently, there has been considerable interest indeﬁning the role of Myc in the biology of a broad range of stemcells. In embryonic stem cells (ESCs), for example, Myc isrequired for maintenance of the pluripotent state ( Cartwrightet al., 2005 ) and for establishment of the stem cell state byway of reprogramming ( Takahashi and Yamanaka, 2006 ).RecentevidencesuggeststhatMyc’sroleinpluripotencyiscon-nectedwithits ability to regulate the cell-cycle machinery. In thisreview, we discuss these recent advances and how an under-standing of Myc and cell-cycle control will lead to a morecomplete understanding of pluripotency and reprogramming.
Pluripotent Cells in Early Embryos: Cell-Cycle Controland Rapid Cell Division
Uncommitted, pluripotent cells in vertebrate pregastrulaembryos undergo extraordinarily rapid rounds of cell division.This trend is associated with cell cycles consisting of alternatingrounds of DNA replication (S phase) and chromosome segrega-tion-cell separation (M phase) without intervening gap phases,such as in cleavage-stage
embryos ( Murray andKirschner, 1989 ). Since all the maternal determinants for devel-opment are deposited in the egg prior to fertilization, zygotictranscription is not required until the midblastula transitionwhen uncommitted cells become speciﬁed. The nature of thiscell-cycle structure thus promotes the rapid ampliﬁcation incell number necessary for early embryonic development. Varia-tionsofthisgeneralthemealsoapplyduringearly
andzebraﬁsh development to enable rapid expansion in cell number just prior to gastrulation ( Edgar and Lehner, 1996; Yarden andGeiger, 1996 ).Inrodents,peri-implantationdevelopmentisdependentonzy-gotic programs, and so cell-cycle progression is driven by denovo RNA and protein synthesis. Expansion of the rodentembryoimmediatelypriortogastrulationisdependentonarapidburst of pluripotent cell proliferation, with average generationtimes estimated to be
4.5–8.0 hr ( Snow, 1977; Lawson et al.,1991 ). These cells exhibit an unusual cell-cycle structure, char-acterized by a short G1 gap phase and a high proportion of Sphase cells (
65%). Since G1 is the period of the cell cyclewhen cell growth primarily occurs, this pattern has often beenused to rationalize why pluripotent cells are small in size. Aspluripotent cells become speciﬁed into one of the three germlayers, the cell cycle typically restructures so that the length of G1isextended,leadingtoincreaseddivisiontimesandageneralincreaseincellvolume( Lawsonetal.,1991 ).Theseobservationsraise the question as to whether there is some mechanistic rela-tionship between pluripotency, cell-cycle structure, and cell sizecontrol.
A Cell-Cycle Structure Synonymous with Pluripotency
65% of their time in S phase and
15% of their time in G1.Upon differentiation, the cell cycle is remodeled such that thepercentage of cells in G1 increases to over 40% and generationtimes increase to >16 hr ( Stead et al., 2002; White et al., 2005 ).These features of cell-cycle control appear to be conserved inrhesus monkey ESCs ( Fluckiger et al., 2006 ), murine epiblaststemcells (EpiSCs;Tesaretal.,2007; Bronsetal.,2007 ), humanand murine induced pluripotent stem cells (iPSCs), and hESCs( Figure 1;Ohtsuka and Dalton, 2008; Dalton, 2009 ). Although
hESCs and human iPSCs exhibit a cell-cycle structure similarto the otherpluripotent celltypes discussed sofar, theircell divi-sioncycletimesaresigniﬁcantlyslower(
32–38hr;OhtsukaandDalton, 2008; Dalton, 2009 ), indicating that rapid division is nota necessaryproperty of pluripotent cells. Thisconclusion issup-ported by other observations showing that slowing the mESCcell cycle with chemical inhibitors has no measurable impacton self-renewal or differentiation potential ( Stead et al., 2002 ).We are unaware of any accurate estimates of generation timesfor human pluripotent cells in early development, and so it is
Cell Stem Cell
, August 7, 2009
2009 Elsevier Inc.