Molecular dissection of telomere dysfunction and analysis of G-overhangs in Arabidopsis thaliana

Telomeres comprise the physical ends of chromosomes. In the absence of telomerase, the enzyme responsible for replenishing telomeric DNA, telomeres progressively shorten due to the end replication problem. Eventually telomeres reach a length where they are recruited into end-to-end chromosome fusions. Through the use of novel PCR strategies, I followed the fate of telomeres in plants lacking telomerase as they progressed into dysfunction. I uncovered two distinct structural/functional length transitions. The first transition (~1 kb) marks the onset of telomere dysfunction, where telomeres are transiently uncapped and a subset of them engage in end-to-end fusions. The second transition (~300 bp) defines complete telomere dysfunction as telomeres below this length lack G-overhangs and the vast majority of the chromosome ends fuse. Thus, these two telomere lengths define architectural transitions that link structure and function. In addition, I uncovered a hierarchy of end-joining pathways that join dysfunctional telomeres in which the non-homologous end-joining (NHEJ) protein, KU predominates. In the absence of KU, telomeres are joined by a microhomologymediated end-joining pathway (MMEJ) that is dependent on Mre11. I also show that DNA ligase IV (LIG4) is the predominant enzyme that ligates dysfunctional telomeres as fusions are reduced in its absence. These studies highlight the importance of repairing DSBs and demonstrate that Arabidopsis possesses highly redundant means for processing dysfunctional telomeres. The G-overhang is an essential feature of the telomere that is required for proper telomere function. I employed methods to examine G-overhang status in various mutants known to contribute to telomere maintenance in Arabidopsis. My analysis revealed that the putative G-overhang binding proteins POT1a, POT1b and POT1c, make modest, but distinct contributions to the G-overhangs. Additionally, I uncovered a major role for the putative telomere capping protein, CIT1 in maintenance of the Goverhang. G-overhang signals obtained from cit1 mutants were grossly increased indicating that CIT1 is involved in either protecting the C-rich strand of the telomere from nuclease attack, or in controlling telomerase extension of the G-strand. Together, these data have provided new insight into factors that contribute to telomere integrity and have further developed Arabidopsis as a model for telomere biology

Telomere dynamics and fusion of critically shortened telomeres in plants lacking DNA ligase IV

In the absence of the telomerase, telomeres undergo progressive shortening and are ultimately recruited into end-to-end chromosome fusions via the non-homologous end joining (NHEJ) double-strand break repair pathway. Previously, we showed that fusion of critically shortened telomeres in Arabidopsis proceeds with approximately the same efficiency in the presence or absence of KU70, a key component of NHEJ. Here we report that DNA ligase IV (LIG4) is also not essential for telomere joining. We observed only a modest decrease (3-fold) in the frequency of chromosome fusions in triple tert ku70 lig4 mutants versus tert ku70 or tert. Sequence analysis revealed that, relative to tert ku70, chromosome fusion junctions in tert ku70 lig4 mutants contained less microhomology and less telomeric DNA. These findings argue that the KU-LIG4 independent end-joining pathway is less efficient and mechanistically distinct from KU-independent NHEJ. Strikingly, in all the genetic backgrounds we tested, chromosome fusions are initiated when the shortest telomere in the population reaches ∼1 kb, implying that this size represents a critical threshold that heralds a detrimental structural transition. These data reveal the transitory nature of telomere stability, and the robust and flexible nature of DNA repair mechanisms elicited by telomere dysfunction

Telomere dynamics and fusion of critically shortened

telomeres in plants lacking DNA ligase I

The Importance of Community Engagement and Research Translation within the NIEHS Superfund Research Program

The National Institute of Environmental Health Sciences Superfund Research Program (SRP) funds university-based, solution-oriented research to understand how hazardous substances contribute to disease and how to prevent exposures to these hazardous substances. A unique aspect of the SRP is that, beyond the biomedical, environmental sciences, and engineering research projects, SRP-funded centers are required to include community engagement to build partnerships with affected communities and research translation to communicate and facilitate the use of research findings. The SRP views both as effective ways to inform and advance science for protection of public health. The purpose of community engagement within the centers is to ensure bidirectional communication between the researchers and the community, identify best practices and activities in community engagement for prevention and intervention activities, enhance knowledge, and support the needs of the communities impacted by hazardous waste sites. The SRP views research translation as communicating and facilitating the use of research findings emanating from the center in a manner most appropriate for their application and for the advancement of a center’s research objectives. The SRP has a strong history of seeking opportunities to work with communities and stakeholders, by translating and sharing research findings in an impactful and informative manner with long-lasting benefits to improve public health

Adapting to Climate Change: Leveraging Systems-Focused Multidisciplinary Research to Promote Resilience

Approximately 2000 official and potential Superfund sites are located within 25 miles of the East or Gulf coasts, many of which will be at risk of flooding as sea levels rise. More than 60 million people across the United States live within 3 miles of a Superfund site. Disentangling multifaceted environmental health problems compounded by climate change requires a multidisciplinary systems approach to inform better strategies to prevent or reduce exposures and protect human health. The purpose of this minireview is to present the National Institute of Environmental Health Sciences Superfund Research Program (SRP) as a useful model of how this systems approach can help overcome the challenges of climate change while providing flexibility to pivot to additional needs as they arise. It also highlights broad-ranging SRP-funded research and tools that can be used to promote health and resilience to climate change in diverse contexts

E-Waste and Harm to Vulnerable Populations: A Growing Global Problem

BACKGROUND: Electronic waste (e-waste) is produced in staggering quantities, estimated globally to be 41.8 million tonnes in 2014. Informal e-waste recycling is a source of much-needed income in many low- to middle-income countries. However, its handling and disposal in underdeveloped countries is often unsafe and leads to contaminated environments. Rudimentary and uncontrolled processing methods often result in substantial harmful chemical exposures among vulnerable populations, including women and children. E-waste hazards have not yet received the attention they deserve in research and public health agendas. OBJECTIVES: We provide an overview of the scale and health risks. We review international efforts concerned with environmental hazards, especially affecting children, as a preface to presenting next steps in addressing health issues stemming from the global e-waste problem. DISCUSSION: The e-waste problem has been building for decades. The increasingly observed adverse health effects from e-waste sites calls for protecting human health and the environment from e-waste contamination. Even if e-waste exposure intervention and prevention efforts are implemented, legacy contamination will remain, necessitating increased awareness of e-waste as a major environmental health threat. CONCLUSION: Global, national, and local levels efforts must aim to create safe recycling operations that consider broad security issues for people who rely on e-waste processing for survival. Paramount to these efforts is reducing pregnant women and children's e-waste exposures to mitigate harmful health effects. With human environmental health in mind, novel dismantling methods and remediation technologies, and intervention practices are needed to protect communities