Polar Growth in the Alphaproteobacterial Order Correction for “Completely Phased Genome Sequencing Through Rhizobiales,” by Pamela J

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Polar Growth in the Alphaproteobacterial Order Correction for “Completely Phased Genome Sequencing Through Rhizobiales,” by Pamela J Corrections MICROBIOLOGY GENETICS, STATISTICS Correction for “Polar growth in the Alphaproteobacterial order Correction for “Completely phased genome sequencing through Rhizobiales,” by Pamela J. B. Brown, Miguel A. de Pedro, David chromosome sorting,” by Hong Yang, Xi Chen, and Wing Hung T. Kysela, Charles Van der Henst, Jinwoo Kim, Xavier De Bolle, Wong, which appeared in issue 1, January 4, 2011, of Proc Natl Clay Fuqua, and Yves V. Brun, which appeared in issue 5, January Acad Sci USA (108:12–17; first published December 15, 2010; 31, 2012, of Proc Natl Acad Sci USA (109:1697–1701; first pub- 10.1073/pnas.1016725108). lished January 17, 2012; 10.1073/pnas.1114476109). The authors note that the database deposition information The authors note that, due to a printer’s error, the affiliation given in the paper is incorrect. Because of privacy concerns, it for David T. Kysela should instead appear as Department of is not possible for the authors to provide the sequence data in Biology, Indiana University, Bloomington, IN 47405. The cor- this study through deposition to a public database. However, the rected author and affiliation lines appear below. The online ver- donor of the DNA sample has given consent for the dataset to sion has been corrected. be shared with qualified researchers under a material transfer agreement. Researchers interested in obtaining this data should Pamela J. B. Browna, Miguel A. de Pedrob, David T. Kyselaa, write to the corresponding author to request the material transfer. Charles Van der Henstc,JinwooKima, Xavier De Bollec, Request for data access should be made to Wing Hung Wong, Clay Fuquaa, and Yves V. Bruna Department of Statistics, Sequoia Hall, 390 Serra Mall, Stanford, CA 94305-4065. Email: [email protected]. aDepartment of Biology, Indiana University, Bloomington, IN 47405; bCentro www.pnas.org/cgi/doi/10.1073/pnas.1200309109 de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas, Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid, Spain; and cResearch Unit in Molecular Biology, Department of Biology, University of Namur, Facultés Universitaires Notre-Dame de la Paix, 5000 Namur, Belgium CHEMISTRY Correction for “Elucidating the mechanism of selective ion ad- www.pnas.org/cgi/doi/10.1073/pnas.1201353109 sorption to the liquid water surface,” by Dale E. Otten, Patrick R. Shaffer, Phillip L. Geissler, and Richard J. Saykally, which appeared in issue 3, January 17, 2012, of Proc Natl Acad Sci USA ECOLOGY (109:701–705; first published January 10, 2012; 10.1073/pnas. Correction for “Dynamic model of flexible phytoplankton nu- 1116169109). trient uptake,” by Juan A. Bonachela, Michael Raghib, and The authors note that, due to a printer’s error, on page 703, Simon A. Levin, which appeared in issue 51, December 20, 2011, left column, first paragraph, lines 9–10, “−6.52 kJ/mol” should of Proc Natl Acad Sci USA (108:20633–20638; first published instead appear as “6.52 kJ/mol.” December 5, 2011; 10.1073/pnas.1118012108). www.pnas.org/cgi/doi/10.1073/pnas.1201349109 The authors note that, due to a printer’s error, on page 20635, right column, first paragraph, line 6 “transporter” should instead appear as “transporters”. Additionally, on page 20635, right column, second para- “A ¼ nðtÞrs =rc ” “A ¼ graph, line 3 rel 2 2 should instead appear as rel 2 2 nðtÞrs =rc ”: Lastly, on page 20636, left column, first full paragraph, line 20, “V ¼ V max ” and second full paragraph, lines 6 and 7, lo should “V ¼ V lo ” instead appear as max . www.pnas.org/cgi/doi/10.1073/pnas.1201165109 3190 | PNAS | February 21, 2012 | vol. 109 | no. 8 www.pnas.org Downloaded by guest on September 30, 2021 Polar growth in the Alphaproteobacterial order Rhizobiales Pamela J. B. Browna, Miguel A. de Pedrob, David T. Kyselac, Charles Van der Henstc, Jinwoo Kima,1, Xavier De Bollec, Clay Fuquaa, and Yves V. Bruna,2 aDepartment of Biology, Indiana University, Bloomington, IN 47405; bCentro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas, Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid, Spain; and cResearch Unit in Molecular Biology, Department of Biology, University of Namur, Facultés Universitaires Notre-Dame de la Paix, 5000 Namur, Belgium Edited* by Eugene W. Nester, University of Washington, Seattle, WA, and approved December 8, 2011 (received for review September 2, 2011) Elongation of many rod-shaped bacteria occurs by peptidoglycan plant pathogen A. tumefaciens as a model bacterium. Time-lapse synthesis at discrete foci along the sidewall of the cells. However, microscopy of individual growing A. tumefaciens cells revealed within the Rhizobiales, there are many budding bacteria, in which that the cell growth was asymmetric and initiation of constriction new cell growth is constrained to a specific region. The phylogeny occurred at the future site of cell division even in relatively small of the Rhizobiales indicates that this mode of zonal growth may cells (Fig. 1B and Movie S1). Transmission electron micrographs be ancestral. We demonstrate that the rod-shaped bacterium showed that the width of the cells at the site of constriction was Agrobacterium tumefaciens grows unidirectionally from the new similar (0.8–0.9 μm) in cells with a daughter cell compartment pole generated after cell division and has an atypical peptidogly- less than 1 μm in length and narrowed as the daughter cell can composition. Polar growth occurs under all conditions tested, compartment length increased further (Fig. 1 C and D). Notably, including when cells are attached to a plant root and under con- a narrow band of FtsZ (Atu_2086) is observed at the site of early ditions that induce virulence. Finally, we show that polar growth constriction in small cells, suggesting that the formation of the also occurs in the closely related bacteria Sinorhizobium meliloti, early constriction may require the assembly of the FtsZ ring (Fig. Brucella abortus, and Ochrobactrum anthropi.Wefind that unipo- 2 A and B). A band of FtsZ is observed at the mid-cell before the lar growth is an ancestral and conserved trait among the Rhizo- initiation of cell division, and a bright focus of FtsZ is found at biales, which includes important mutualists and pathogens of the mid-cell of deeply constricted cells (Fig. 2 A). A small focus MICROBIOLOGY plants and animals. of FtsZ persists at the new cell pole for a short period after cell division; however, the polar focus disappears before the forma- cell wall morphogenesis | cell elongation-division cycle tion of the band of FtsZ at the site of early constriction. The position of FtsZ in the constriction appears to remain fixed longation of most rod-shaped bacterial cells is thought be relative to the old pole as the cell grows, suggesting that new cell Emediated by peptidoglycan synthesis along the sidewall of the growth is largely constrained to the daughter cell compartment cells and depends on the elongase, a protein complex consisting (Fig. 2B and Movie S2). Plotting the position of the constriction of MreBCD, RodA, and PBP2 [reviewed (1)]. Peptidoglycan (as observed in the transmission electron micrographs) relative synthesis continues laterally until a cell has roughly doubled in to the cell poles indicates that the constriction shifts slightly away length, at which point new cell wall precursors are directed to the from the mother cell pole as the cell nears division (Fig. S1A). division site by the bacterial tubulin homolog FtsZ (2). After Tracking the length of the mother cell compartment over time in completion of cell division, peptidoglycan biosynthesis is redir- dividing cells reveals that there is only a small increase in mother ected to lateral growth along the sidewall. cell length after cell division (Fig. S1 B and C). These data Although these modes of cell growth are generally thought to suggest that the size of the mother cell compartment is relatively be predominant among rod-shaped bacteria, in Actinobacteria, constant and we infer that new cell growth occurs primarily elongation occurs at the cell poles and requires the polarity de- within the daughter cell compartment. terminant protein DivIVA (3–8). In addition, a striking type of To identify regions of new cell growth, A. tumefaciens cells polar growth is found within some Alphaproteobacteria that were pulse labeled with the amine reactive dye Texas red-X grow by budding (9) (Fig. 1A). In budding bacteria, new daughter succinimidyl ester (TRSE), which stains cell surface proteins. In E. coli cells emerge strictly from the pole of the mother cell. Thus, we the signal from TRSE-stained proteins dissipated homo- presume that new cell wall synthesis must be restricted to a small genously along the sidewalls of the cells as a consequence of polar area. The scattered distribution of budding bacteria within lateral cell growth as previously shown (21) but remained trap- the Rhizobiales (Fig. 1A) suggests an ancestral origin of budding, ped at the old cell poles, which are comprised of inert peptido- A fl indicating that members of the Rhizobiaceae and Brucellaceae glycan (Fig. 3 and Movie S3). In contrast, the uorescent signal fi A. tumefaciens B C families may also grow by polar extension rather than their from TRSE remained xed in (Fig. 3 and , B usually assumed dispersed mode of growth. Further support for Fig. S2 , and Movies S4, S5, and S6), suggesting that the main this hypothesis is provided by the observation that mutations or body of the cells, and not just the poles, were inert. When the treatments that impair cell division in Brucella abortus (10, 11), Sinorhizobium meliloti (12, 13), and Agrobacterium tumefaciens (14–18) cause branching morphologies to arise, rather than fil- Author contributions: P.J.B.B., M.A.d.P., D.T.K., C.F., and Y.V.B.
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