Characterization of the mIF4G Domains in the RNA Surveillance Protein Upf2p.

Thirty percent of all mutations causing human disease generate mRNAs with premature termination codons (PTCs). Recognition and degradation of these PTC-containing mRNAs is carried out by the mechanism known as nonsense-mediated mRNA decay (NMD). Upf2 is a scaffold protein known to be a central component of the NMD surveillance pathway. It harbors three middle domains of eukaryotic initiation factor 4G (mIF4G-1, mIF4G-2, mIF4G-3) in its N-terminal region that are potentially important in regulating the surveillance pathway. In this study, we defined regions within the mIF4G-1 and mIF4G-2 that are required for proper function of Upf2p in NMD and translation termination in Saccharomyces cerevisiae. In addition, we narrowed down the activity of these regions to an aspartic acid (D59) in mIF4G-1 that is important for NMD activity and translation termination accuracy. Taken together, these studies suggest that inherently charged residues within mIF4G-1 of Upf2p play a role in the regulation of the NMD surveillance mechanism in S. cerevisiae.

Ten simple rules for running a summer research program.

To continue to advance the field of computational biology and fill the constantly growing need for new trainees who are well positioned for success, immersive summer research experiences have proven to be effective in preparing students to navigate the challenges that lay ahead in becoming future computational biologists. Here, we describe 10 simple rules for planning, offering, running, and improving a summer research program in computational biology that supports students in honing technical competencies for success in research and developing skills to become successful scientific professionals.

CREARE: A Course-Based Undergraduate Research Experience To Study the Responses of the Endangered Coral Acropora cervicornis to a Changing Environment.

There is mounting evidence to support that students who participate in scientific research experiences are more likely to continue on to advanced degrees and careers in science, technology, engineering, and mathematics (STEM). To introduce more students to the benefits of research, we have drawn on an ongoing project aimed at understanding how the Caribbean staghorn coral Acropora cervicornis responds to environmental fluctuations to develop a semester-long course-based undergraduate research experience (CURE), entitled CREARE (Coral Response to Environment Authentic Research Experience). The main mode of instruction in CREARE is through topic modules, and course evaluation is achieved through writing assignments. Students in CREARE perform experiments in the laboratory to measure the abundance of photo-protective proteins in coral tissue from samples collected at different depths and at different times of the year and analyze environmental data using the R programming language. CREARE participants have contributed to the progress of the research project by generating novel data and making improvements to experimental protocols. Furthermore, pre- and post-course assessment of content knowledge revealed that students perform significantly better on a written exam after participating in CREARE, while also displaying appreciable shifts in attitudes towards science in student perception surveys. In addition, through qualitative analysis of focus group interviews, we gathered evidence to suggest that mediating variables that predict students' persistence in science are bolstered through our application of the CURE modality. Overall, CREARE can serve as a model for developing more research-based courses that successfully engage students in scientific research.

The microbial biosphere of the coral Acropora cervicornis in Northeastern Puerto Rico.

BACKGROUND: Coral reefs are the most biodiverse ecosystems in the marine realm, and they not only contribute a plethora of ecosystem services to other marine organisms, but they also are beneficial to humankind via, for instance, their role as nurseries for commercially important fish species. Corals are considered holobionts (host + symbionts) since they are composed not only of coral polyps, but also algae, other microbial eukaryotes and prokaryotes. In recent years, Caribbean reef corals, including the once-common scleractinian coral Acropora cervicornis, have suffered unprecedented mortality due to climate change-related stressors. Unfortunately, our basic knowledge of the molecular ecophysiology of reef corals, particularly with respect to their complex bacterial microbiota, is currently too poor to project how climate change will affect this species. For instance, we do not know how light influences microbial communities of A. cervicornis, arguably the most endangered of all Caribbean coral species. To this end, we characterized the microbiota of A. cervicornis inhabiting water depths with different light regimes. METHODS: Six A. cervicornis fragments from different individuals were collected at two different depths (three at 1.5 m and three at 11 m) from a reef 3.2 km off the northeastern coast of Puerto Rico. We characterized the microbial communities by sequencing the 16S rRNA gene region V4 with the Illumina platform. RESULTS: A total of 173,137 good-quality sequences were binned into 803 OTUs with a 97% similarity. We uncovered eight bacterial phyla at both depths with a dominance of 725 Rickettsiales OTUs (Proteobacteria). A fewer number (38) of low dominance OTUs varied by depth and taxa enriched in shallow water corals included Proteobacteria (e.g. Rhodobacteraceae and Serratia) and Firmicutes (Streptococcus). Those enriched in deeper water corals featured different Proteobacterial taxa (Campylobacterales and Bradyrhizobium) and Firmicutes (Lactobacillus). DISCUSSION: Our results confirm that the microbiota of A. cervicornis inhabiting the northeastern region of Puerto Rico is dominated by a Rickettsiales-like bacterium and that there are significant changes in less dominant taxa at different water depths. These changes in less dominant taxa may potentially impact the coral's physiology, particularly with respect to its ability to respond to future increases in temperature and CO2.

CtIP interacts with TopBP1 and Nbs1 in the response to double-stranded DNA breaks (DSBs) in Xenopus egg extracts.

In the presence of double-stranded DNA breaks (DSBs), the activation of ATR is achieved by the ability of ATM to phosphorylate TopBP1 on serine 1131, which leads to an enhancement of the interaction between ATR and TopBP1. In Xenopus egg extracts, the Mre11-Rad50-Nbs1 (MRN) complex is additionally required to bridge ATM and TopBP1 together. In this report, we show that CtIP, which is recruited to DSB-containing chromatin, interacts with both TopBP1 and Nbs1 in a damage-dependent manner. An N-terminal region containing the first two BRCT repeats of TopBP1 is essential for the interaction with CtIP. Furthermore, two distinct regions in the N-terminus of CtIP participate in establishing the association between CtIP and TopBP1. The first region includes two adjacent putative ATM/ATR phosphorylation sites on serines 273 and 275. Secondly, binding is diminished when an MRN-binding region spanning residues 25-48 is deleted, indicative of a role for the MRN complex in mediating this interaction. This was further evidenced by a decrease in the interaction between CtIP and TopBP1 in Nbs1-depleted extracts and a reciprocal decrease in the binding of Nbs1 to TopBP1 in the absence of CtIP, suggestive of the formation of a complex containing CtIP, TopBP1, and the MRN complex. When CtIP is immunodepleted from egg extracts, the activation of the response to DSBs is compromised and the levels of ATR, TopBP1, and Nbs1 on damaged chromatin are reduced. Thus, CtIP interacts with TopBP1 in a damage-stimulated, MRN-dependent manner during the activation of ATR in response to DSBs.