Role of the Srs2-Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae.

Saccharomyces cerevisiae Srs2, in addition to its well-documented antirecombination activity, has been proposed to play a role in promoting synthesis-dependent strand annealing (SDSA). Here we report the identification and characterization of an SRS2 mutant with a single amino acid substitution (srs2-F891A) that specifically affects the Srs2 pro-SDSA function. This residue is located within the Srs2-Rad51 interaction domain and embedded within a protein sequence resembling a BRC repeat motif. The srs2-F891A mutation leads to a complete loss of interaction with Rad51 as measured through yeast two-hybrid analysis and a partial loss of interaction as determined through protein pull-down assays with purified Srs2, Srs2-F891A, and Rad51 proteins. Even though previous work has shown that internal deletions of the Srs2-Rad51 interaction domain block Srs2 antirecombination activity in vitro, the Srs2-F891A mutant protein, despite its weakened interaction with Rad51, exhibits no measurable defect in antirecombination activity in vitro or in vivo Surprisingly, srs2-F891A shows a robust shift from noncrossover to crossover repair products in a plasmid-based gap repair assay, but not in an ectopic physical recombination assay. Our findings suggest that the Srs2 C-terminal Rad51 interaction domain is more complex than previously thought, containing multiple interaction sites with unique effects on Srs2 activity

Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase δ-extending D-loops.

Synthesis-dependent strand annealing (SDSA) is the preferred mode of homologous recombination in somatic cells leading to an obligatory non-crossover outcome, thus avoiding the potential for chromosomal rearrangements and loss of heterozygosity. Genetic analysis identified the Srs2 helicase as a prime candidate to promote SDSA. Here, we demonstrate that Srs2 disrupts D-loops in an ATP-dependent fashion and with a distinct polarity. Specifically, we partly reconstitute the SDSA pathway using Rad51, Rad54, RPA, RFC, DNA Polymerase δ with different forms of PCNA. Consistent with genetic data showing the requirement for SUMO and PCNA binding for the SDSA role of Srs2, Srs2 displays a slight but significant preference to disrupt extending D-loops over unextended D-loops when SUMOylated PCNA is present, compared to unmodified PCNA or monoubiquitinated PCNA. Our data establish a biochemical mechanism for the role of Srs2 in crossover suppression by promoting SDSA through disruption of extended D-loops

A Randomized Trial Examining Preoperative Sedative Medication and Post-operative Sleep in Children

Study Objective Midazolam has been found to have beneficial effects on anxiety in children in the preoperative setting. Prior studies have examined various postoperative behaviors of children, but little research has examined the effects of preoperative use of midazolam with postoperative sleep. The purpose of this investigation was to compare postoperative sleep in children as a function of preoperative sedative medication use. Design This study was a 2-group randomized controlled trial. Setting Participants were recruited from Yale-New Haven Children\u27s Hospital. Patients Participants included a convenience sample of 70 children between the ages of 3 to 12 years undergoing ambulatory tonsillectomy and adenoidectomy. Interventions Children were randomly assigned to 1 of 2 groups: a control group who received preoperative acetaminophen only (n = 32) and an experimental group who received both acetaminophen and midazolam preoperatively (n = 38). Measurements Parents completed measures of postoperative behavioral recovery and a subset of children wore actigraphs to examine objective sleep data. Main Results Children who received midazolam experienced similar sleep changes compared to children in the control group. The actigraph data revealed that children who received midazolam were awake significantly less during the night compared to the control group (P= .01). Conclusion Children who received midazolam before surgery had similar postoperative sleep changes compared to children who did not receive midazolam. Further understanding of the postoperative behavioral effects of midazolam on children will help guide healthcare providers in their practice

The COS Absorption Survey of Baryon Harbors (CASBaH): Warm-hot Circumgalactic Gas Reservoirs Traced by Ne VIII Absorption

We survey the highly ionized circumgalactic media (CGM) of 29 blindly selected galaxies at 0.49 < z_(gal) < 1.44 based on high-S/N ultraviolet spectra of z > 1 QSOs and the galaxy database from the COS Absorption Survey of Baryon Harbors (CASBaH). We detect the Ne VIII doublet in nine of the galaxies, and for gas with N(Ne VIII) > 10^13.3 cm^-2 (> 10^13.5 cm^-2), we derive a Ne VIII covering fraction f_c = 75 +15/-25% (44 +22/-20%) within impact parameter (rho) < 200 kpc of M_* = 10^(9.5-11.5) Msol galaxies and f_c = 70 +16/-22% (f_c = 42 +20/-17%) within rho < 1.5 virial radii. We estimate the mass in Ne VIII-traced gas to be M_gas(Ne VIII) > 10^9.5 Msol (Z/Zsol)^-1, or 6-20% of the expected baryonic mass if the Ne VIII absorbers have solar metallicity. Ionizing Ne VII to Ne VIII requires 207 eV, and photons with this energy are scarce in the CGM. However, for the median halo mass and redshift of our sample, the virial temperature is close to the peak temperature for the Ne VIII ion, and the Ne VIII-bearing gas is plausibly collisionally ionized near this temperature. Moreover, we find that photoionized Ne VIII requires cool and low-density clouds that would be highly underpressured (by approximately two orders of magnitude) relative to the putative, ambient virialized medium, complicating scenarios where such clouds could survive. Thus, more complex (e.g., non-equilibrium) models may be required; this first statistical sample of Ne VIII absorber/galaxy systems will provide stringent constraints for future CGM studies.Comment: Published in ApJL, Volume 877, Issue 2, Article L2

TESS Data Release Notes: Sector 18 DR25

This release note discusses the science data products produced by the Science Processing Operations Center at Ames Research Center from Sector 18 observations made with the TESS spacecraft and cameras as a means to document instrument performance and data characteristics

TESS Data Release Notes: Sector 17, DR24

This release note discusses the science data products produced by the Science Processing Operations Center at Ames Research Center from Sector 17 observations made with the TESS spacecraft and cameras as a means to document instrument performance and data characteristics

TESS Data Release Notes: Sector 16, DR22

This release note discusses the science data products produced by the Science Processing Operations Center at Ames Research Center from Sector 16 observations made with the TESS spacecraft and cameras as a means to document instrument performance and data characteristics

TESS Data Release Notes: Sector 20, DR27

This release note discusses the science data products produced by the Science Processing Operations Center at Ames Research Center from Sector 20 observations made with the TESS spacecraft and cameras as a means to document instrument performance and data characteristics

TESS Data Release Notes: Sector 9 DR11

This release note discusses the science data products produced by the Science Processing Operations Center at Ames Research Center from Sector 9 observations made with the TESS spacecraft and cameras as a means to document instrument performance and data characteristics