Biology subjects

Murakami, H.

Publications and source records attributed to Murakami, H..

25 records · Page 2Linked to original sources

bioRxiv2020

Mutual anticipation can contribute to self-organization in human crowds

Human crowds provide paradigmatic examples of collective behavior emerging through self-organization. Although the underlying interaction has been considered to obey the distance-dependent law, resembling physical particle systems, recent findings emphasized that pedestrian motions are fundamentally influenced by the anticipated future positions of their neighbors rather than their current positions. Therefore, anticipatory interaction may play a crucial role in collective patterning. However, whether and how individual anticipation functionally benefits the group is not well-understood. We suggest that collective patterning in human crowds is promoted by anticipatory path-seeking behavior resulting in a scale-free movement pattern, called the Levy walk. In our experiments of lane formation, a striking example of self-organized patterning in human crowds where people moving in opposite directions spontaneously segregate into several unidirectional lanes, we manipulated some pedestrians ability to anticipate by having them type on a mobile phone while walking. The manipulation slowed overall walking speeds and delayed the onset of global patterning, and the distracted pedestrians sometimes failed to achieve their usual walking strategy. Moreover, we observed that the delay of global patterning depends on decisions made by pedestrians who were moving toward the distracted ones and had no choice but to take sudden large steps, presumably because of difficulty in anticipating the motions of their counterparts. These results imply that mutual anticipation between pedestrians facilitates efficient transition to emergent patterning in situations where nobody within a crowd is distracted. Our findings may contribute to efficient crowd management and inform future models of self-organizing systems.

animal behavior and cognition↗

bioRxiv2020

Chromosome-autonomous feedback downregulates meiotic DSB competence upon synaptonemal complex formation

The number of DNA double-strand breaks (DSBs) initiating meiotic recombination is elevated in Saccharomyces cerevisiae mutants that are globally defective in forming crossovers and synaptonemal complex (SC), a protein scaffold juxtaposing homologous chromosomes. These mutants thus appear to lack a negative feedback loop that inhibits DSB formation when homologs engage one another. This feedback is predicted to be chromosome autonomous, but this has not been tested. Moreover, what chromosomal process is recognized as "homolog engagement" remains unclear. To address these questions, we evaluated effects of homolog engagement defects restricted to small portions of the genome using karyotypically abnormal yeast strains with a homeologous chromosome V pair, monosomic V, or trisomy XV. We found that homolog-engagement-defective chromosomes incurred more DSBs, concomitant with prolonged retention of the DSB-promoting protein Rec114, while the rest of the genome remained unaffected. SC-deficient, crossover-proficient mutants ecm11 and gmc2 experienced increased DSB numbers diagnostic of homolog engagement defects. These findings support the hypothesis that SC formation provokes DSB protein dissociation, leading in turn to loss of a DSB competent state. Our findings show that DSB number is regulated in a chromosome-autonomous fashion and provide insight into how homeostatic DSB controls respond to aneuploidy during meiosis. bioRxiv version 2 (September 2020) We added one bioRxiv citation in the discussion section which shows increased breaks in gmc2 and ecm11 mutants. This work was done independently by Keun Kim, Miki Shinohara and colleagues. The pulse gel electrophoresis result shown in Figure 1D has been re-quantified. The conclusion remains that homeologous and monosomic chromosome V generate more DSBs compared to internally controlled chromosome III. One zip3 map included in PCA (Figure 6C) and hierarchical clustering analysis (Figure S5B) has been deleted because this particular map has not yet been published. This change does not affect the conclusion. An additional supplemental table (S5) has been added to show Spo11-oligo datasets used in this study. Figures and preprint have been reformatted. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=193 SRC="FIGDIR/small/089367v2_fig1.gif" ALT="Figure 1"> View larger version (67K): org.highwire.dtl.DTLVardef@871eaforg.highwire.dtl.DTLVardef@ef7995org.highwire.dtl.DTLVardef@176441org.highwire.dtl.DTLVardef@1b6e772_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 1.C_FLOATNO Higher DSB levels on homolog-engagement defective chromosomes. (A) Cartoons of wild-type, homeologous, monosomic and trisomic chromosome configurations. Grey lines are S. cerevisiae chromosomes and orange lines are S. pastorianus. The homeologous chrV pair rarely synapses or recombines. The trisomic chromosomes can adopt different synaptic configurations ("II+I", partner switch, and triple synapsis). (B) Representative PFGE Southern blots probed for chrIII and chrV. P, signal from parental-length DNA; W, signal in wells. Asterisk indicates an ectopic recombination product between leu2::hisG (on chrIII) and ho::hisG (on chrIV) in the monosomic strain. The other strains do not form this product because they do not have a hisG insert at ho. (C) Poisson-corrected DSB quantification of PFGE Southern blots. (D) Quantification of time-averaged DSBs on chrV relative to chrIII for wild-type, homeologous and monosomic strains. * p<0.05, unpaired t test. Error bars in C, D are mean {+/-} SD except for the monosomic strain (mean {+/-} range). C_FIG O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=167 SRC="FIGDIR/small/089367v2_fig6.gif" ALT="Figure 6"> View larger version (36K): org.highwire.dtl.DTLVardef@1152edforg.highwire.dtl.DTLVardef@1b853daorg.highwire.dtl.DTLVardef@89392dorg.highwire.dtl.DTLVardef@108d65a_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 6.C_FLOATNO gmc2{Delta} and ecm11{Delta} mutants share similar homolog-engagement signatures as zip3{Delta}. (A, B) Representative labeling of Spo11-oligo complexes in gmc2{Delta} (A) and ecm11{Delta} (B) mutants and quantification relative to wild-type cultures processed in parallel. Error bars indicate mean {+/-} SD for 3 cultures. Radiolabeled Spo11-oligo complexes are detected by autoradiography (left panel, top) and total Spo11 is detected by anti-Flag western blot (WB; left panel, bottom). The two main labeled species differ in the sizes of oligos (Neale et al. 2005). Most Spo11 protein does not end up making DSBs, so nearly all of the visible western blot signal is from free Spo11 that does not have an oligo attached (Neale et al. 2005). (C) Principal component analysis of 21 wild-type and mutant Spo11-oligo maps. (D) Loss of anticorrelation between chromosome length and DSB density in homolog-engagement-defective mutants. Each point is one chromosome. Correlation coefficients (Pearsons r) are shown. (E) Fold change of Spo11-oligo counts in different chromosomal domains. Tel, within 20 kb of telomeres; Cen, within 10 kb of centromeres; rDNA, from 60 kb leftward to 30 kb rightward of rDNA; Interstitial, all others. Horizontal dashed lines mark values assumed as no change (black) and average change (1.8-fold for zip3{Delta}, cyan; 1.7-fold for ecm11{Delta}, purple). Boxes indicate median and interquartile range; whiskers indicate the most extreme data points that are [≤] 1.5 times the interquartile range from the box; individual points are outliers C_FIG O_FIG O_LINKSMALLFIG WIDTH=174 HEIGHT=200 SRC="FIGDIR/small/089367v2_figS5.gif" ALT="Figure 5"> View larger version (35K): org.highwire.dtl.DTLVardef@130cbf6org.highwire.dtl.DTLVardef@9ebe8forg.highwire.dtl.DTLVardef@1f0a2b4org.highwire.dtl.DTLVardef@14a0fcf_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOSupplemental Figure S5.C_FLOATNO DSB patterns in gmc2{Delta} and ecm11{Delta} mutants. (A) Meiotic progression showing percentage of cells completing the first divisions (total bi- and tetra-nucleate cells, mean {+/-} SD, 3 cultures each). At least 100 cells were counted at each time point for each culture. (B) Hierarchical clustering analysis using the "Ward D2" method for 21 wild-type and mutant maps. The y-axis represents height of the cluster dendrogram. In the label for each data set, the letters A, B, C and D stand for maps generated by Xiaojing Mu, Neeman Mohibullah, Megan van Overbeek, and Xuan Zhu, respectively, and the numerals distinguish biological replicate maps. (C) Regional variation in response to zip3{Delta} and ecm11{Delta} mutations for chromosome XII. Lines, local regression (loess) based on Spo11-oligo density changes in 5 kb-bins; green circle, centromere; rDNA location is labeled. (D, E) Fold change of Spo11-oligo densities for zip3{Delta} and ecm11{Delta} compared to wild type in pericentric (D) and telomere-proximal (E) regions. Lines, local regression (loess) for data in 5 kb-bins averaged across 32 chromosome arms; dashed line, no change in black and genome average change in cyan (1.8-fold, zip3{Delta}) and purple (1.7-fold, ecm11{Delta}); yellow shading, zones suppressed for DSB formation as previously defined (Pan etal. 2011). C_FIG O_TBL View this table: org.highwire.dtl.DTLVardef@15408baorg.highwire.dtl.DTLVardef@9171aorg.highwire.dtl.DTLVardef@cbabfaorg.highwire.dtl.DTLVardef@187d594org.highwire.dtl.DTLVardef@d6eb68_HPS_FORMAT_FIGEXP M_TBL O_FLOATNOSupplemental Table S5.C_FLOATNO O_TABLECAPTIONSpo11-oligo datasets used in this study C_TABLECAPTION C_TBL

genetics↗

bioRxiv2018

Estimating fish population abundance by integrating quantitative data on environmental DNA and hydrodynamic modelling

We propose a general framework of abundance estimation based on spatially replicated quantitative measurements of environmental DNA in which production, transport, and degradation of DNA are explicitly accounted for. Application to a Japanese jack mackerel (Trachurus japonicus) population in Maizuru Bay revealed that the method gives an estimate of population abundance comparable to that of a quantitative echo sounder method. These findings indicate the ability of environmental DNA to reliably reflect population abundance of aquatic macroorganisms and may o[ff]er a new avenue for population monitoring based on the fast, cost-e[ff]ective, and non-invasive sampling of genetic information.

ecology↗

bioRxiv2018

Multilayered mechanisms ensure that short chromosomes recombine in meiosis

To segregate accurately during meiosis, homologous chromosomes in most species must recombine. Very small chromosomes would risk missegregation if recombination were randomly distributed, so the double-strand breaks (DSBs) that initiate recombination are not haphazard. How this nonrandomness is controlled is not under-stood. Here we demonstrate that Saccharomyces cerevisiae integrates multiple, temporally distinct pathways to regulate chromosomal binding of pro-DSB factors Rec114 and Mer2, thereby controlling duration of a DSB-competent state. Homologous chromosome engagement regulates Rec114/Mer2 dissociation late in prophase, whereas replication timing and proximity to centromeres or telomeres influence timing and amount of Rec114/Mer2 accumulation early. A distinct early mechanism boosts Rec114/Mer2 binding quickly to high levels specifically on the shortest chromosomes, dependent on chromosome axis proteins and subject to selection pressure to maintain hyperrecombinogenic properties of these chromosomes. Thus, an organisms karyotype and its attendant risk of meiotic missegregation influence the shape and evolution of its recombination landscape.

genetics↗

bioRxiv2018

Potential Application of Bacteriophages in Enrichment Culture for Improved Prenatal Streptococcus agalactiae Screening

Vertical transmission of Streptococcus agalactiae can cause neonatal infections. A culture test in the late stage of pregnancy is used to screen for the presence of maternal S. agalactiae for intrapartum antibiotic prophylaxis. For the test, vaginal-rectal swab sampling is immediately followed by enrichment culture and bacterial identification. In some cases, Enterococcus faecalis competes with and overgrowths S. agalactiae in the enrichment culture. Consequently, the identification test occasionally yields false-negative results. Bacterial viruses, bacteriophages (phages), infect and kill specific host bacteria. In the current study, we explored the feasibility of using phages to minimize the undesirable E. faecalis outgrowth and facilitate S. agalactiae detection in an experimental setting. Phage mixture was prepared using three phages that specifically infect E. faecalis: phiEF24C, phiEF17H, and phiM1EF22. The mixture inhibited the growth of 86.7% (26/30) of E. faecalis strains tested in the enrichment broth. When single strains of E. faecalis and S. agalactiae were inoculated in the enrichment broth containing the phage mixture, bacterial growth was inhibited or facilitated, respectively. Further, several sets of S. agalactiae and E. faecalis strains were co-cultured, and bacteria were detected on chromogenic agar after the enrichment culture. S. agalactiae was dominant after plating a phage mixture-treated co-culture, while it was barely detected after plating the untreated co-culture. Considering these observations, the phage mixture can be employed in the S. agalactiae culture test to increase test accuracy.

microbiology↗

bioRxiv2017

Quantitative monitoring of multispecies fish environmental DNA using high-throughput sequencing

Effective ecosystem conservation and resource management require quantitative monitoring of biodiversity, including accurate descriptions of species composition and temporal variations of species abundance. Therefore, quantitative monitoring of biodiversity has been performed for many ecosystems, but it is often time- and effort-consuming and costly. Recent studies have shown that environmental DNA (eDNA), which is released to the environment from macro-organisms living in a habitat, contains information about species identity and abundance. Thus, analyzing eDNA would be a promising approach for more efficient biodiversity monitoring. In the present study, we added internal standard DNAs (i.e., known amounts of short DNA fragments from fish species that have never been observed in a sampling area) to eDNA samples, which were collected weekly from a coastal marine ecosystem in Maizuru-Bay, Kyoto, Japan (from April 2015 to March 2016), and performed metabarcoding analysis using Illumina MiSeq to simultaneously identify fish species and quantify fish eDNA copy numbers. A correction equation was obtained for each sample using the relationship between the number of sequence reads and the added amount of the standard DNAs, and this equation was used to estimate the copy numbers from the sequence reads of non-standard fish eDNA. The calculated copy numbers showed significant positive correlation with those determined by quantitative PCR, suggesting that eDNA metabarcoding with standard DNA enabled useful quantification of eDNA. Furthermore, for samples that show a high level of PCR inhibition, our method might allow more accurate quantification than qPCR because the correction equations generated using internal standard DNAs would include the effect of PCR inhibition. A single run of Illumina MiSeq produced > 70 quantitative fish eDNA time series in our study, showing that our method could contribute to more efficient and quantitative monitoring of biodiversity.

ecology↗

bioRxiv2017

Histone H3 threonine 11 phosphorylation is catalyzed directly by the meiosis-specific kinase Mek1 and provides a molecular readout of Mek1 activity in vivo

Saccharomyces cerevisiae Mek1 is a CHK2/Rad53-family kinase that regulates meiotic recombination and progression upon its activation in response to DNA double-strand breaks (DSBs). The full catalog of direct Mek1 phosphorylation targets remains unknown. Here, we show that phosphorylation of histone H3 on threonine 11 (H3 T11ph) is induced by meiotic DSBs in S. cerevisiae and Schizosaccharomyces pombe. Molecular genetic experiments in S. cerevisiae confirmed that Mek1 is required for H3 T11ph and revealed that phosphorylation is rapidly reversed when Mek1 kinase is no longer active. Reconstituting histone phosphorylation in vitro with recombinant proteins demonstrated that Mek1 directly catalyzes H3 T11 phosphorylation. Mutating H3 T11 to nonphosphorylatable residues conferred no detectable defects in otherwise unperturbed meiosis, although the mutations modestly reduced spore viability in certain strains where Rad51 is used for strand exchange in place of Dmc1. H3 T11ph is therefore mostly dispensable for Mek1 function. However, H3 T11ph provides an excellent marker of ongoing Mek1 kinase activity in vivo. Anti-H3 T11ph chromatin immunoprecipitation followed by deep sequencing demonstrated that H3 T11ph was highly enriched at presumed sites of attachment of chromatin to chromosome axes, gave a more modest signal along chromatin loops, and was present at still lower levels immediately adjacent to DSB hotspots. These localization patterns closely tracked the distribution of Red1 and Hop1, axis proteins required for Mek1 activation. These findings provide insight into the spatial disposition of Mek1 kinase activity and the higher order organization of recombining meiotic chromosomes.\n\nbioRxiv version 2 (June 2017)One major experimental change was incorporated into the revised manuscript: We repeated the anti-H3 T11ph ChIP-seq experiment on larger scale, including two meiotic time points from each of two wild type cultures and one time point from a spo11-Y135F mutant culture. To facilitate comparison of different samples, we used meiotic S. pombe cells as a spike-in control for all samples for both anti-H3 and anti-H3 T11ph ChIP-seq. Most conclusions described in the first bioRxiv submission were confirmed, but the improved datasets allowed us to derive more detailed information in particular about H3 T11ph patterns around DSB sites.\n\nbioRxiv version 3 (October 2017)The following experimental changes were incorporated, along with more minor changes in response to reviewer comments: O_LIWe added previously unpublished ChIP-seq data for Red1 protein, generated by Masaru Ito and Kunihiro Ohta, who have been added as coauthors.\nC_LIO_LIWe repeated key experiments with the H3-T11V single point mutant. No conclusions were changed relative to prior experiments with the H3-S10, T11V mutant.\nC_LIO_LIWe repeated the analysis of spore viability in a dmc1 rad54-T132A background using a more appropriate isogenic control, and recapitulated the original conclusion that the H3-T11V mutation modestly decreases spore viability in this sensitized background.\nC_LI

molecular biology↗