The JCMT 12CO(3-2) Survey of the Cygnus X Region: I. A Pathfinder

Cygnus X is one of the most complex areas in the sky. This complicates interpretation, but also creates the opportunity to investigate accretion into molecular clouds and many subsequent stages of star formation, all within one small field of view. Understanding large complexes like Cygnus X is the key to understanding the dominant role that massive star complexes play in galaxies across the Universe. The main goal of this study is to establish feasibility of a high-resolution CO survey of the entire Cygnus X region by observing part of it as a Pathfinder, and to evaluate the survey as a tool for investigating the star-formation process. A 2x4 degree area of the Cygnus X region has been mapped in the 12CO(3-2) line at an angular resolution of 15" and a velocity resolution of ~0.4km/s using HARP-B and ACSIS on the James Clerk Maxwell Telescope. The star formation process is heavily connected to the life-cycle of the molecular material in the interstellar medium. The high critical density of the 12CO(3-2) transition reveals clouds in key stages of molecule formation, and shows processes that turn a molecular cloud into a star. We observed ~15% of Cygnus X, and demonstrated that a full survey would be feasible and rewarding. We detected three distinct layers of 12CO(3-2) emission, related to the Cygnus Rift (500-800 pc), to W75N (1-1.8 kpc), and to DR21 (1.5-2.5 kpc). Within the Cygnus Rift, HI self-absorption features are tightly correlated with faint diffuse CO emission, while HISA features in the DR21 layer are mostly unrelated to any CO emission. 47 molecular outflows were detected in the Pathfinder, 27 of them previously unknown. Sequentially triggered star formation is a widespread phenomenon.Comment: 18 pages, 13 figures, accepted for publication in Astronomy & Astrophysic

Targeting the Replication Initiator of the Second Vibrio Chromosome: Towards Generation of Vibrionaceae-Specific Antimicrobial Agents

The Vibrionaceae is comprised of numerous aquatic species and includes several human pathogens, such as Vibrio cholerae, the cause of cholera. All organisms in this family have two chromosomes, and replication of the smaller one depends on rctB, a gene that is restricted to the Vibrionaceae. Given the increasing prevalence of multi-drug resistance in pathogenic vibrios, there is a need for new targets and drugs to combat these pathogens. Here, we carried out a high throughput cell-based screen to find small molecule inhibitors of RctB. We identified a compound that blocked growth of an E. coli strain bearing an rctB-dependent plasmid but did not influence growth of E. coli lacking this plasmid. This compound, designated vibrepin, had potent cidal activity against V. cholerae and inhibited the growth of all vibrio species tested. Vibrepin blocked RctB oriCII unwinding, apparently by promoting formation of large non-functional RctB complexes. Although vibrepin also appears to have targets other than RctB, our findings suggest that RctB is an attractive target for generation of novel antibiotics that only block growth of vibrios. Vibrio-specific agents, unlike antibiotics currently used in clinical practice, will not engender resistance in the normal human flora or in non-vibrio environmental microorganisms

Fragmentation and mass segregation in the massive dense cores of Cygnus X

We present Plateau de Bure interferometer observations obtained in continuum at 1.3 and 3.5 mm towards the six most massive and young (IR-quiet) dense cores in Cygnus X. Located at only 1.7 kpc, the Cygnus X region offers the opportunity of reaching small enough scales (of the order of 1700 AU at 1.3 mm) to separate individual collapsing objects. The cores are sub-fragmented with a total of 23 fragments inside 5 cores. Only the most compact core, CygX-N63, could actually be a single massive protostar with an envelope mass as large as 60 Msun. The fragments in the other cores have sizes and separations similar to low-mass pre-stellar and proto-stellar condensations in nearby protoclusters, and are probably of the same nature. A total of 9 out of these 23 protostellar objects are found to be probable precursors of OB stars with envelope masses ranging from 6 to 23 Msun. The level of fragmentation is globally higher than in the turbulence regulated, monolithic collapse scenario, but is not as high as expected in a pure gravo-turbulent scenario where the distribution of mass is dominated by low-mass protostars/stars. Here, the fractions of the total core masses in the high-mass fragments are reaching values as high as 28, 44, and 100 % in CygX-N12, CygX-N53, and CygX-N63, respectively, much higher than what an IMF-like mass distribution would predict. The increase of the fragmentation efficiency as a function of density in the cores is proposed to be due to the increasing importance of self-gravity leading to gravitational collapse at the scale of the dense cores. At the same time, the cores tend to fragment into a few massive protostars within their central regions. We are therefore probably witnessing here the primordial mass segregation of clusters in formation.Comment: 14 pages, 16 figures, submitted for publication in A&

Regulatory Cross-Talk Links Vibrio cholerae Chromosome II Replication and Segregation

There is little knowledge of factors and mechanisms for coordinating bacterial chromosome replication and segregation. Previous studies have revealed that genes (and their products) that surround the origin of replication (oriCII) of Vibrio cholerae chromosome II (chrII) are critical for controlling the replication and segregation of this chromosome. rctB, which flanks one side of oriCII, encodes a protein that initiates chrII replication; rctA, which flanks the other side of oriCII, inhibits rctB activity. The chrII parAB2 operon, which is essential for chrII partitioning, is located immediately downstream of rctA. Here, we explored how rctA exerts negative control over chrII replication. Our observations suggest that RctB has at least two DNA binding domains—one for binding to oriCII and initiating replication and the other for binding to rctA and thereby inhibiting RctB's ability to initiate replication. Notably, the inhibitory effect of rctA could be alleviated by binding of ParB2 to a centromere-like parS site within rctA. Furthermore, by binding to rctA, ParB2 and RctB inversely regulate expression of the parAB2 genes. Together, our findings suggest that fluctuations in binding of the partitioning protein ParB2 and the chrII initiator RctB to rctA underlie a regulatory network controlling both oriCII firing and the production of the essential chrII partitioning proteins. Thus, by binding both RctB and ParB2, rctA serves as a nexus for regulatory cross-talk coordinating chrII replication and segregation

DNA Adenine Methylation Is Required to Replicate Both Vibrio cholerae Chromosomes Once per Cell Cycle

DNA adenine methylation is widely used to control many DNA transactions, including replication. In Escherichia coli, methylation serves to silence newly synthesized (hemimethylated) sister origins. SeqA, a protein that binds to hemimethylated DNA, mediates the silencing, and this is necessary to restrict replication to once per cell cycle. The methylation, however, is not essential for replication initiation per se but appeared so when the origins (oriI and oriII) of the two Vibrio cholerae chromosomes were used to drive plasmid replication in E. coli. Here we show that, as in the case of E. coli, methylation is not essential for oriI when it drives chromosomal replication and is needed for once-per-cell-cycle replication in a SeqA-dependent fashion. We found that oriII also needs SeqA for once-per-cell-cycle replication and, additionally, full methylation for efficient initiator binding. The requirement for initiator binding might suffice to make methylation an essential function in V. cholerae. The structure of oriII suggests that it originated from a plasmid, but unlike plasmids, oriII makes use of methylation for once-per-cell-cycle replication, the norm for chromosomal but not plasmid replication

FtsK-Dependent Dimer Resolution on Multiple Chromosomes in the Pathogen Vibrio cholerae

Unlike most bacteria, Vibrio cholerae harbors two distinct, nonhomologous circular chromosomes (chromosome I and II). Many features of chromosome II are plasmid-like, which raised questions concerning its chromosomal nature. Plasmid replication and segregation are generally not coordinated with the bacterial cell cycle, further calling into question the mechanisms ensuring the synchronous management of chromosome I and II. Maintenance of circular replicons requires the resolution of dimers created by homologous recombination events. In Escherichia coli, chromosome dimers are resolved by the addition of a crossover at a specific site, dif, by two tyrosine recombinases, XerC and XerD. The process is coordinated with cell division through the activity of a DNA translocase, FtsK. Many E. coli plasmids also use XerCD for dimer resolution. However, the process is FtsK-independent. The two chromosomes of the V. cholerae N16961 strain carry divergent dimer resolution sites, dif1 and dif2. Here, we show that V. cholerae FtsK controls the addition of a crossover at dif1 and dif2 by a common pair of Xer recombinases. In addition, we show that specific DNA motifs dictate its orientation of translocation, the distribution of these motifs on chromosome I and chromosome II supporting the idea that FtsK translocation serves to bring together the resolution sites carried by a dimer at the time of cell division. Taken together, these results suggest that the same FtsK-dependent mechanism coordinates dimer resolution with cell division for each of the two V. cholerae chromosomes. Chromosome II dimer resolution thus stands as a bona fide chromosomal process