View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Current Biology 22, R340–R349, May 8, 2012 ª2012 Elsevier Ltd All rights reserved DOI 10.1016/j.cub.2012.02.032 Cell Size Control in Bacteria Review An-Chun Chien, Norbert S. Hill, and Petra Anne Levin exhibit a vast array of morphologies ranging from rods and filaments to cocci, spirals and amoeboid-like forms. The diversity of bacteria is mirrored in the size of individual Like eukaryotes, bacteria must coordinate division with species, which range from w0.3 mm for obligate intracellular growth to ensure cells are the appropriate size for a pathogenic members of the genus Mycoplasma,tow600 mm given environmental condition or developmental fate. As for Epulopiscium fishelsoni, a Gram-positive commensal single-celled organisms, nutrient availability is one of the inhabitant of Surgeonfish guts, and 750 mm for Thiomargarita strongest influences on bacterial cell size. Classic physio- namibiensis, a chemilithotrophic Gram-negative bacterium logical experiments conducted over four decades ago native to coastal Namibia [1–3]. While T. namibiensis is first demonstrated that cell size is directly correlated essentially a large gas vesicle surrounded by a thin layer of with nutrient source and growth rate in the Gram-negative cytoplasm, Epulopiscium has managed to overcome diffu- bacterium Salmonella typhimurium. This observation sub- sion-dependent limitations on cell size, in part by increasing sequently served as the basis for studies revealing a role genome number along with cell size. These tens-of-thou- for cell size in cell cycle progression in a closely related sands of genomes are arranged around the periphery of organism, Escherichia coli. More recently, the develop- the Epulopiscium cell, where they are thought to facilitate ment of powerful genetic, molecular, and imaging tools responses to local stimuli and thereby contribute to mainte- has allowed us to identify and characterize the nutrient- nance of the extremely large cell size [4]. dependent pathway responsible for coordinating cell divi- Similarly, although it is not the focus of this review, it is sion and cell size with growth rate in the Gram-positive important to note that cell size and shape are, not surpris- model organism Bacillus subtilis. Here, we discuss the ingly, sensitive to changes in the morphogenesis of the role of cell size in bacterial growth and development and bacterial cell wall. In particular, enzymes involved in synthe- propose a broadly applicable model for cell size control sizing the peptidoglycan material that constitutes the bacte- in this important and highly divergent domain of life. rial cell wall, as well as the Mre proteins which recent data suggest help coordinate peptidoglycan synthesis, all play Introduction an important role in cell size control by maintaining cell Coordinating growth with division is essential to ensure that shape and width within normal parameters. For excellent cells are the appropriate size for a given environmental reviews on this topic see [5–7]. condition or developmental fate. This is true not only for multicellular plants and animals, but also for single-celled Binary Fission: A Deceptively Simple Mode organisms that need to adapt quickly to rapid changes in of Reproduction environmental conditions. Like their eukaryotic counter- Bacteria exhibit many forms of reproduction, including parts, in the absence of environmental or internal pressure binary fission, budding (Planctomycetes), hyphal growth to increase size, exponentially growing bacteria cultured (Actinomycetes), daughter cell formation (Epulopiscium), under a constant set of parameters exhibit little size varia- and the formation of multicellular baeocytes (the cyanobac- tion between cells. Maintenance of cell size within a narrow terium Stanieria). Of these, binary fission is one of the most band indicates that cells have mechanisms to transiently common, and is by far the best understood. adjust the timing of division to correct aberrations in Binary fission in B. subtilis and E. coli is deceptively cell size generated through stochastic events. Similarly, simple. During exponential growth, cells double in mass although bacterial cell size is essentially constant under and then divide in the middle to produce equivalently sized steady state conditions, environmental challenges and de- daughter cells. Despite its apparent simplicity, binary fission velopmental programs frequently require changes in cell is in fact the culmination of a complex, elaborately orches- size just as they do for other single-celled and multicellular trated series of events. Binary fission requires cells to double organisms. in mass, initiate and terminate at least one round of chromo- Here we review what is known about bacterial cell size some replication, decatenate and segregate sister chromo- control during steady state growth, in response to changes somes (also referred to as nucleoids), assemble the in nutrient availability, and during development. For reasons division machinery precisely at mid-cell, and coordinate of brevity we will focus on two well-studied model systems, membrane invagination with cell wall synthesis to form Escherichia coli and Bacillus subtilis. Where appropriate a complete septum (Figure 1). we will also include information gleaned from work in other In contrast to eukaryotes, the bacterial cell cycle is not organisms. divided into discrete stages. Instead cell growth, DNA repli- Before we delve into the discussion of bacterial cell size cation, chromosome segregation, and even the initial control in mesophilic model systems, it is important to note assembly of the division machinery can overlap with one that bacteria occupy habitats that include thermal vents another, a physically challenging proposition at faster where temperatures are well over 100C, 5 M saline salt growth rates. Because of the overlapping processes, the pools, and environments where ionizing radiation levels are nomenclature used for describing stages of the eukaryotic a thousand times the lethal dose for humans. Bacteria also cell cycle (G1, S, G2, and M) is not useful when describing the bacterial cell cycle. The alternative nomenclature includes three discrete periods: B, the time between cell Department of Biology, Box 1137, Washington University, 1 Brookings birth and the initiation of DNA replication; C, the period Dr., Saint Louis, MO, USA. required for chromosome replication; and D, the time E-mail: [email protected] between the termination of replication and division. Special Issue R341 Figure 1. The bacterial division cycle. AB FtsZ assembly is coordinated with DNA Cell division Initiation of replication replication and segregation. (A) (1) In new- born cells, FtsZ (red) is unassembled. A 5 1 circular chromosome (blue) with a single origin of replication (green) is located at mid-cell. (2–4) After chromosome replica- tion initiates, the origins of replication Termination Elongation and and origin separation separate and move to opposite poles of nucleoid 4 2 the cell. Once replication is complete, the segregation condensed chromosomes separate, leaving 3 a nucleoid free space. (3) FtsZ ring forma- tion coincides with chromosome segrega- tion. Assembly starts from a single point Assembly of FtsZ ring Current Biology at mid-cell and extends bidirectionally. (5) During cytokinesis, the FtsZ ring constricts at the leading edge of the invaginating membrane. (B) Immunofluorescence micrograph of expo- nentially growing B. subtilis cells with FtsZ rings. Arrows indicate examples of cells with rings. Bar = 5 mm. Under steady state conditions, B. subtilis and E. coli initiated per division cycle [16,24]. The ratio of free cells exhibit little variation in average cell size beyond the DnaAATP to oriC then increases in a growth-rate-dependent requirements of binary fission [8,9]. Maintaining cell size manner until it is high enough to support initiation, a point within these parameters suggests cells precisely control that is coincident with achievement of a specific mass in both the timing and position of cell division and can compen- wild-type E. coli [16,24]. sate for stochastic events that lead to a reduction in cell size Consistent with a role for DnaA in the size-dependent or an increase in cell size by transiently altering the length of regulation of replication initiation, significantly reducing their cell cycle (Figure 2). Although changes in the duration of DnaA expression delays initiation and increases cell size, any cell cycle event can theoretically impact cell size, in while overexpressing DnaA leads to premature initiation E. coli and B. subtilis, only two, the initiation of DNA replica- and a reduction in cell size in both E. coli and B. subtilis tion and cell division, have been implicated as important [14,23]. Similarly, short E. coli mutants delay initiation until control points in the homeostatic regulation of cell size. they reach a size that is approximately equivalent to wild- Below we discuss the role of initiation and division in the type cells [25]. This delay is alleviated following a modest spatial and temporal control of cell size under steady state increase in DnaA expression, supporting the idea that conditions. growth-dependent accumulation of active DnaA to critical levels is the primary trigger for initiation in E. coli. Cell Size and the Initiation of DNA Replication Although it is easy to imagine how normalizing size at