An extreme paucity of second population AGB stars in the normal globular cluster M4

Galactic Globular clusters (GCs) are now known to harbour multiple stellar populations, which are chemically distinct in many light element abundances. It is becoming increasingly clear that asymptotic giant branch (AGB) stars in GCs show different abundance distributions in light elements compared to those in the red giant branch (RGB) and other phases, skewing toward more primordial, field-star-like abundances, which we refer to as subpopulation one (SP1). As part of a larger program targeting giants in GCs, we obtained high-resolution spectra for a sample of 106 RGB and 15 AGB stars in Messier 4 (NGC 6121) using the 2dF+HERMES facility on the Anglo-Australian Telescope. In this Letter we report an extreme paucity of AGB stars with [Na/O] > -0.17 in M4, which contrasts with the RGB that has abundances up to [Na/O] =0.55. The AGB abundance distribution is consistent with all AGB stars being from SP1. This result appears to imply that all subpopulation two stars (SP2; Na-rich, O-poor) avoid the AGB phase. This is an unexpected result given M4's horizontal branch morphology -- it does not have an extended blue horizontal branch. This is the first abundance study to be performed utilising the HERMES spectrograph.Comment: 5 pages, 2 figures, 4 tables (full Table 1 online). Accepted for publication in MNRAS Letter

AGB subpopulations in the nearby globular cluster NGC 6397

It has been well established that Galactic Globular clusters (GCs) harbour more than one stellar population, distinguishable by the anti-correlations of light element abundances (C-N, Na-O, and Mg-Al). These studies have been extended recently to the asymptotic giant branch (AGB). Here we investigate the AGB of NGC 6397 for the first time. We have performed an abundance analysis of high-resolution spectra of 47 RGB and 8 AGB stars, deriving Fe, Na, O, Mg and Al abundances. We find that NGC 6397 shows no evidence of a deficit in Na-rich AGB stars, as reported for some other GCs - the subpopulation ratios of the AGB and RGB in NGC 6397 are identical, within uncertainties. This agrees with expectations from stellar theory. This GC acts as a control for our earlier work on the AGB of M 4 (with contrasting results), since the same tools and methods were used.Comment: 10 pages, 7 figures, 8 tables (2 online-only). Accepted for publication in MNRA

On the AGB stars of M 4: A robust disagreement between spectroscopic observations and theory

Several recent spectroscopic investigations have presented conflicting results on the existence of Na-rich asymptotic giant branch (AGB) stars in the Galactic globular cluster M4 (NGC6121). The studies disagree on whether or not Na-rich red giant branch (RGB) stars evolve to the AGB. For a sample of previously published HER- MES/AAT AGB and RGB stellar spectra we present a re-analysis of O, Na, and Fe abundances, and a new analysis of Mg and Al abundances; we also present CN band strengths for this sample, derived from low-resolution AAOmega spectra. Following a detailed literature comparison, we find that the AGB samples of all studies consistently show lower abundances of Na and Al, and are weaker in CN, than RGB stars in the cluster. This is similar to recent observations of AGB stars in NGC 6752 and M 62. In an attempt to explain this result, we present new theoretical stellar evolutionary models for M 4; however, these predict that all stars, including Na-rich RGB stars, evolve onto the AGB. We test the robustness of our abundance results using a variety of atmospheric models and spectroscopic methods; however, we do not find evidence that systematic modelling uncertainties can explain the apparent lack of Na- rich AGB stars in M4. We conclude that an unexplained, but robust, discordance between observations and theory remains for the AGB stars in M 4.Comment: 25 pages, 21 figures, 15 tables, accepted for publication in MNRA

PCNA stimulates catalysis by structure-specific nucleases using two distinct mechanisms: substrate targeting and catalytic step

The sliding clamp Proliferating Cell Nuclear Antigen (PCNA) functions as a recruiter and organizer of a wide variety of DNA modifying enzymes including nucleases, helicases, polymerases and glycosylases. The 5′-flap endonuclease Fen-1 is essential for Okazaki fragment processing in eukaryotes and archaea, and is targeted to the replication fork by PCNA. Crenarchaeal XPF, a 3′-flap endonuclease, is also stimulated by PCNA in vitro. Using a novel continuous fluorimetric assay, we demonstrate that PCNA activates these two nucleases by fundamentally different mechanisms. PCNA stimulates Fen-1 by increasing the enzyme's binding affinity for substrates, as suggested previously. However, PCNA activates XPF by increasing the catalytic rate constant by four orders of magnitude without affecting the KM. PCNA may function as a platform upon which XPF exerts force to distort DNA substrates, destabilizing the substrate and/or stabilizing the transition state structure. This suggests that PCNA can function directly in supporting catalysis as an essential cofactor in some circumstances, a new role for a protein that is generally assumed to perform a passive targeting and organizing function in molecular biology. This could provide a mechanism for the exquisite control of nuclease activity targeted to specific circumstances, such as replication forks or damaged DNA with pre-loaded PCNA

Biochemical Characterization of a Structure-Specific Resolving Enzyme from Sulfolobus islandicus Rod-Shaped Virus 2

Sulfolobus islandicus rod shaped virus 2 (SIRV2) infects the archaeon Sulfolobus islandicus at extreme temperature (70°C–80°C) and acidity (pH 3). SIRV2 encodes a Holliday junction resolving enzyme (SIRV2 Hjr) that has been proposed as a key enzyme in SIRV2 genome replication. The molecular mechanism for SIRV2 Hjr four-way junction cleavage bias, minimal requirements for four-way junction cleavage, and substrate specificity were determined. SIRV2 Hjr cleaves four-way DNA junctions with a preference for cleavage of exchange strand pairs, in contrast to host-derived resolving enzymes, suggesting fundamental differences in substrate recognition and cleavage among closely related Sulfolobus resolving enzymes. Unlike other viral resolving enzymes, such as T4 endonuclease VII or T7 endonuclease I, that cleave branched DNA replication intermediates, SIRV2 Hjr cleavage is specific to four-way DNA junctions and inactive on other branched DNA molecules. In addition, a specific interaction was detected between SIRV2 Hjr and the SIRV2 virion body coat protein (SIRV2gp26). Based on this observation, a model is proposed linking SIRV2 Hjr genome resolution to viral particle assembly

Reversion of a fungal genetic code alteration links proteome instability with genomic and phenotypic diversification

Many fungi restructured their proteomes through incorporation of serine (Ser) at thousands of protein sites coded by the leucine (Leu) CUG codon. How these fungi survived this potentially lethal genetic code alteration and its relevance for their biology are not understood. Interestingly, the human pathogen Candida albicans maintains variable Ser and Leu incorporation levels at CUG sites, suggesting that this atypical codon assignment flexibility provided an effective mechanism to alter the genetic code. To test this hypothesis, we have engineered C. albicans strains to misincorporate increasing levels of Leu at protein CUG sites. Tolerance to the misincorporations was very high, and one strain accommodated the complete reversion of CUG identity from Ser back to Leu. Increasing levels of Leu misincorporation decreased growth rate, but production of phenotypic diversity on a phenotypic array probing various metabolic networks, drug resistance, and host immune cell responses was impressive. Genome resequencing revealed an increasing number of genotype changes at polymorphic sites compared with the control strain, and 80% of Leu misincorporation resulted in complete loss of heterozygosity in a large region of chromosome V. The data unveil unanticipated links between gene translational fidelity, proteome instability and variability, genome diversification, and adaptive phenotypic diversity. They also explain the high heterozygosity of the C. albicans genome and open the door to produce microorganisms with genetic code alterations for basic and applied research.We thank Alexander Johnson for providing the C. albicans strains and plasmids and Judith Berman, Csaba Pál, and Dieter Söll for their useful comments and suggestions on the manuscript. The study was funded by the European Union Framework Program 7 (EUFP7) Sybaris Consortium Project 242220 and the Portuguese Science Foundation through Fundo Europeu de Desenvolvimento Regional (FEDER/FCT) Project PTDC/BIA-MIC/099826/2008.publishe