The spatial organization of replication is determined by cell size independently of chromosome copy number

2021 ◽  
Author(s):  
Anna Knoppel ◽  
Oscar Brostrom ◽  
Konrad Gras ◽  
David Fange ◽  
Johan Elf
Keyword(s):  
Cell Size ◽  
Growth Rates ◽  
Spatial Organization ◽  
Fixed Number ◽  
Replication Forks ◽  
Genetic Perturbations ◽  
Switch Mechanism ◽  
Marker Frequency ◽  
Rule Out ◽  
As If

We have studied how Escherichia coli coordinates replication and division cycles by tracking replisomes in individual cells through thousands of division cycles. By analysing the replication and division of the cells at various growth rates and with different genetic perturbations, we have reached four major conclusions: (1) Initiation is not based on titration of DnaA to the chromosomal binding sites because initiation is still coordinated with the division cycle several generations after DnaA synthesis is turned off. (2) We can rule out that initiation is triggered by a simple DnaA-ADP/ATP switch mechanism because initiation is still triggered at a well-defined cell size in a DARS1/DARS2/datA triple knockout if DnaA is overexpressed. (3) From the spatial organization of replisomes at different growth rates, it appears as if initiation is triggered by termination, but this hypothesis is ruled out by an NGS-based marker frequency analysis. (4) Overlapping replication cycles do not affect the spatial organization of the replication machinery and it appears as if each replication center contains a fixed number of DnaN molecules independently of how many replication forks are being processed.

scholarly journals Threshold effect of growth rate on population variability of Escherichia coli cell lengths

2017 ◽  
Vol 4 (2) ◽  
pp. 160417 ◽  
Author(s):  
Manasi S. Gangan ◽  
Chaitanya A. Athale
Keyword(s):  
Escherichia Coli ◽  
Growth Rate ◽  
Cell Size ◽  
Growth Rates ◽  
Threshold Effect ◽  
Cell Length ◽  
Replication Forks ◽  
E Coli ◽  
Cell Size Distributions ◽  
Rate Threshold

A long-standing question in biology is the effect of growth on cell size. Here, we estimate the effect of Escherichia coli growth rate ( r ) on population cell size distributions by estimating the coefficient of variation of cell lengths (CV L ) from image analysis of fixed cells in DIC microscopy. We find that the CV L is constant at growth rates less than one division per hour, whereas above this threshold, CV L increases with an increase in the growth rate. We hypothesize that stochastic inhibition of cell division owing to replication stalling by a RecA-dependent mechanism, combined with the growth rate threshold of multi-fork replication (according to Cooper and Helmstetter), could form the basis of such a threshold effect. We proceed to test our hypothesis by increasing the frequency of stochastic stalling of replication forks with hydroxyurea (HU) treatment and find that cell length variability increases only when the growth rate exceeds this threshold. The population effect is also reproduced in single-cell studies using agar-pad cultures and ‘mother machine’-based experiments to achieve synchrony. To test the role of RecA, critical for the repair of stalled replication forks, we examine the CV L of E. coli ΔrecA cells. We find cell length variability in the mutant to be greater than wild-type, a phenotype that is rescued by plasmid-based RecA expression. Additionally, we find that RecA-GFP protein recruitment to nucleoids is more frequent at growth rates exceeding the growth rate threshold and is further enhanced on HU treatment. Thus, we find growth rates greater than a threshold result in increased E. coli cell lengths in the population, and this effect is, at least in part, mediated by RecA recruitment to the nucleoid and stochastic inhibition of division.

Klinefelter's Syndrome with Atypical Presenting Features

1981 ◽  
Vol 26 (8) ◽  
pp. 567-568 ◽  
Author(s):  
V.K. Yeragani ◽  
G. Hopkinson
Keyword(s):  
Personality Disorder ◽  
Temporal Lobe Epilepsy ◽  
Temporal Lobe ◽  
Klinefelter’S Syndrome ◽  
Klinefelter's Syndrome ◽  
Abnormal Behaviour ◽  
Hearing Voices ◽  
Rule Out ◽  
As If

The authors thought it would be of interest to present this case which proved to be Klinefelter's syndrome with “xxy” pattern, as the patient presented with episodes of abnormal behaviour lasting for a few minutes. He would suddenly become anxious, look perplexed, start clenching his fists and at times complain of thoughts that some persons were trying to attack him. He also felt as if he were hearing voices talking to him. Observation in the hospital and EEG recording helped to rule out temporal lobe epilepsy and a diagnosis of Klinefelter's syndrome with personality disorder and psychogenic attacks of abnormal behaviour was made at the time of discharge.

Combining Probability Forecasts: 60% and 60% Is 60%, but Likely and Likely Is Very Likely

2021 ◽  
Author(s):  
Robert Mislavsky ◽  
Celia Gaertig
Keyword(s):  
Decision Analysis ◽  
Combination Strategy ◽  
Consumer Decisions ◽  
Probability Forecasts ◽  
Rule Out ◽  
As If

How do we combine others’ probability forecasts? Prior research has shown that when advisors provide numeric probability forecasts, people typically average them (i.e., they move closer to the average advisor’s forecast). However, what if the advisors say that an event is “likely” or “probable?” In eight studies (n = 7,334), we find that people are more likely to act as if they “count” verbal probabilities (i.e., they move closer to certainty than any individual advisor’s forecast) than they are to “count” numeric probabilities. For example, when the advisors both say an event is “likely,” participants will say that it is “very likely.” This effect occurs for both probabilities above and below 50%, for hypothetical scenarios and real events, and when presenting the others’ forecasts simultaneously or sequentially. We also show that this combination strategy carries over to subsequent consumer decisions that rely on advisors’ likelihood judgments. We discuss and rule out several candidate mechanisms for our effect. This paper was accepted by Yuval Rottenstreich, decision analysis.

scholarly journals Rapid Growth of UropathogenicEscherichia coliduring Human Urinary Tract Infection

mBio ◽  
2018 ◽  
Vol 9 (2) ◽  
Author(s):  
Valerie S. Forsyth ◽  
Chelsie E. Armbruster ◽  
Sara N. Smith ◽  
Ali Pirani ◽  
A. Cody Springman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Growth Rate ◽  
Urinary Tract ◽  
Growth Rates ◽  
High Growth ◽  
Replication Forks ◽  
Chromosomal Replication ◽  
E Coli ◽  
Tract Infections

ABSTRACTUropathogenicEscherichia coli(UPEC) strains cause most uncomplicated urinary tract infections (UTIs). These strains are a subgroup of extraintestinal pathogenicE. coli(ExPEC) strains that infect extraintestinal sites, including urinary tract, meninges, bloodstream, lungs, and surgical sites. Here, we hypothesize that UPEC isolates adapt to and grow more rapidly within the urinary tract than otherE. coliisolates and survive in that niche. To date, there has not been a reliable method available to measure their growth ratein vivo. Here we used two methods: segregation of nonreplicating plasmid pGTR902, and peak-to-trough ratio (PTR), a sequencing-based method that enumerates bacterial chromosomal replication forks present during cell division. In the murine model of UTI, UPEC strain growth was robustin vivo, matching or exceedingin vitrogrowth rates and only slowing after reaching high CFU counts at 24 and 30 h postinoculation (hpi). In contrast, asymptomatic bacteriuria (ABU) strains tended to maintain high growth ratesin vivoat 6, 24, and 30 hpi, and population densities did not increase, suggesting that host responses or elimination limited population growth. Fecal strains displayed moderate growth rates at 6 hpi but did not survive to later times. By PTR,E. coliin urine of human patients with UTIs displayed extraordinarily rapid growth during active infection, with a mean doubling time of 22.4 min. Thus, in addition to traditional virulence determinants, including adhesins, toxins, iron acquisition, and motility, very high growth ratesin vivoand resistance to the innate immune response appear to be critical phenotypes of UPEC strains.IMPORTANCEUropathogenicEscherichia coli(UPEC) strains cause most urinary tract infections in otherwise healthy women. While we understand numerous virulence factors are utilized byE. colito colonize and persist within the urinary tract, these properties are inconsequential unless bacteria can divide rapidly and survive the host immune response. To determine the contribution of growth rate to successful colonization and persistence, we employed two methods: one involving the segregation of a nonreplicating plasmid in bacteria as they divide and the peak-to-trough ratio, a sequencing-based method that enumerates chromosomal replication forks present during cell division. We found that UPEC strains divide extraordinarily rapidly during human UTIs. These techniques will be broadly applicable to measurein vivogrowth rates of other bacterial pathogens during host colonization.

scholarly journals Cell size sensing in animal cells coordinates anabolic growth rates and cell cycle progression to maintain cell size uniformity

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Miriam Bracha Ginzberg ◽  
Nancy Chang ◽  
Heather D'Souza ◽  
Nish Patel ◽  
Ran Kafri ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Size ◽  
Growth Rates ◽  
Cell Cycle Progression ◽  
Cellular Growth ◽  
Small Cells ◽  
Live Cells ◽  
Control Mechanisms ◽  
Animal Cells ◽  
Size Uniformity

Cell size uniformity in healthy tissues suggests that control mechanisms might coordinate cell growth and division. We derived a method to assay whether cellular growth rates depend on cell size, by monitoring how variance in size changes as cells grow. Our data revealed that, twice during the cell cycle, growth rates are selectively increased in small cells and reduced in large cells, ensuring cell size uniformity. This regulation was also observed directly by monitoring nuclear growth in live cells. We also detected cell-size-dependent adjustments of G1 length, which further reduce variability. Combining our assays with chemical/genetic perturbations confirmed that cells employ two strategies, adjusting both cell cycle length and growth rate, to maintain the appropriate size. Additionally, although Rb signaling is not required for these regulatory behaviors, perturbing Cdk4 activity still influences cell size, suggesting that the Cdk4 pathway may play a role in designating the cell’s target size.

scholarly journals Co-arrays in the Next Fortran Standard

2007 ◽  
Vol 15 (1) ◽  
pp. 9-26 ◽  
Author(s):  
John Reid ◽  
Robert W. Numrich
Keyword(s):  
Fixed Number ◽  
Fortran Program ◽  
Local Data ◽  
Current Image ◽  
Data Objects ◽  
Access To Data ◽  
Array Syntax ◽  
As If

The WG5 committee, at its meeting in Delft, May 2005, decided to include co-arrays in the next Fortran Standard. A Fortran program containing co-arrays is interpreted as if it were replicated a fixed number of times and all copies were executed asynchronously. Each copy has its own set of data objects and is called an image. The array syntax of Fortran is extended with additional trailing subscripts in square brackets to give a clear and straightforward representation of access to data on other images. References without square brackets are to local data, so code that can run independently is uncluttered. Any occurrence of square brackets is a warning about communication between images. The additional syntax requires support in the compiler, but it has been designed to be easy to implement and to give the compiler scope both to apply its optimizations within each image and to optimize the communication between images. The extension includes execution control statements for synchronizing images and intrinsic procedures to return the number of images, to return the index of the current image, and to perform collective operations. The paper does not attempt to describe the full details of the feature as it now appears in the draft of the new standard. Instead, we describe a subset and demonstrate the use of this subset with examples.

scholarly journals Picoplankton Diel Variability and Estimated Growth Rates in Epipelagic and Mesopelagic Waters of the Central Red Sea

2021 ◽  
Vol 8 ◽  
Author(s):  
Najwa Al-Otaibi ◽  
Francisca C. García ◽  
Xosé Anxelu G. Morán
Keyword(s):  
Cell Size ◽  
Red Sea ◽  
Growth Rates ◽  
Specific Growth ◽  
Nucleic Acid Content ◽  
Autotrophic Picoplankton ◽  
Heterotrophic Prokaryotes ◽  
Diel Variability ◽  
Specific Growth Rates

The diel variability of the abundance and cell size of picoplanktonic groups in the central Red Sea was monitored every 2 h in situ on 4 occasions (once per season) from 2015 to 2016. We distinguished Prochlorococcus, low (LF-Syn) and high (HF-Syn) fluorescence Synechococcus, small (Speuk) and large (Lpeuk) picoeukaryotes and two groups of heterotrophic prokaryotes of low (LNA) and high (HNA) nucleic acid content. The diel variability in abundance was less marked than in cell size and more apparent in autotrophs than heterotrophs. Specific growth rates were estimated by an empirical relationship from measurements obtained in bottle incubations of surface and deep samples collected in the winter compared with in situ variations in cell size over 24 h. Autotrophic picoplankton groups generally grew faster (0.23–0.77 d–1) than heterotrophic prokaryotes (0.12–0.50 d–1). Surface to 100 m depth-weighted specific growth rates displayed a clear seasonal pattern for Prochlorococcus, with maxima in winter (0.77 ± 0.07 d–1) and minima in fall (0.52 ± 0.07 d–1). The two groups of Synechococcus peaked in spring, with slightly higher growth rates of LF-Syn (0.57 ± 0.04 d–1) than HF-Syn (0.43 ± 0.04 d–1). Speuk and Lpeuk showed different seasonal patterns, with lower values of the former (0.27 ± 0.02 and 0.37 ± 0.04 d–1, respectively). HNA consistently outgrew LNA heterotrophic prokaryotes, with a higher growth in the epipelagic (0–200 m, 0.36 ± 0.03 d–1) than in the mesopelagic (200–700 m, 0.26 ± 0.03 d–1), while no differences were found for LNA cells (0.19 ± 0.03 d–1 and 0.17 ± 0.02 d–1, respectively). With all data pooled, the mean diel abundances of autotrophic picoplankton in the upper epipelagic and of HNA cells in the epipelagic and mesopelagic layers were significantly correlated with the specific growth rates estimated from cell size variations. Our high-resolution sampling dataset suggests that changes in growth rates underlie the noticeable seasonality of picoplankton recently described in these tropical waters.

scholarly journals Replication Fork Velocities at Adjacent Replication Origins Are Coordinately Modified during DNA Replication in Human Cells

2007 ◽  
Vol 18 (8) ◽  
pp. 3059-3067 ◽  
Author(s):  
Chiara Conti ◽  
Barbara Saccà ◽  
John Herrick ◽  
Claude Lalou ◽  
Yves Pommier ◽  
...  
Keyword(s):  
Dna Replication ◽  
Single Molecule ◽  
Replication Fork ◽  
Spatial Organization ◽  
Genome Duplication ◽  
Functional Organization ◽  
Replication Forks ◽  
Dynamic Regulation ◽  
Timely Completion ◽  
Genome Level

The spatial organization of replicons into clusters is believed to be of critical importance for genome duplication in higher eukaryotes, but its functional organization still remains to be fully clarified. The coordinated activation of origins is insufficient on its own to account for a timely completion of genome duplication when interorigin distances vary significantly and fork velocities are constant. Mechanisms coordinating origin distribution with fork progression are still poorly elucidated, because of technical difficulties of visualizing the process. Taking advantage of a single molecule approach, we delineated and compared the DNA replication kinetics at the genome level in human normal primary and malignant cells. Our results show that replication forks moving from one origin, as well as from neighboring origins, tend to exhibit the same velocity, although the plasticity of the replication program allows for their adaptation to variable interorigin distances. We also found that forks that emanated from closely spaced origins tended to move slower than those associated with long replicons. Taken together, our results indicate a functional role for origin clustering in the dynamic regulation of genome duplication.

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