Autonomous Plasmid-like Replication of a Conjugative Transposon

Integrative and conjugative elements (ICEs), a.k.a. conjugative transposons, are mobile genetic elements involved in many biological processes, including pathogenesis, symbiosis and the spread of antibiotic resistance. Unlike conjugative plasmids that are extra-chromosomal and replicate autonomously, ICEs are integrated in the chromosome and replicate passively during chromosomal replication. It is generally thought that ICEs do not replicate autonomously. We found that when induced, Bacillus subtilis ICEBs1 undergoes autonomous plasmid-like replication. Replication was unidirectional, initiated from the ICEBs1 origin of transfer, oriT, and required the ICEBs1-encoded relaxase NicK. Replication also required several host proteins needed for chromosomal replication, but did not require the replicative helicase DnaC or the helicase loader protein DnaB. Rather, replication of ICEBs1 required the helicase PcrA that is required for rolling circle replication of many plasmids. Transfer of ICEBs1 from the donor required PcrA, but did not require replication, indicating that PcrA, and not DNA replication, facilitates unwinding of ICEBs1 DNA for horizontal transfer. Although not needed for horizontal transfer, replication of ICEBs1 was needed for stability of the element. We propose that autonomous plasmid-like replication is a common property of ICEs and contributes to the stability and maintenance of these mobile genetic elements in bacterial populations.National Institutes of Health (U.S.) (grant GM50895

The Bacillus subtilis conjugative transposon ICEBs1 mobilizes plasmids lacking dedicated mobilization functions

Integrative and conjugative elements (ICEs, also known as conjugative transposons) are mobile elements that are found integrated in a host genome and can excise and transfer to recipient cells via conjugation. ICEs and conjugative plasmids are found in many bacteria and are important agents of horizontal gene transfer and microbial evolution. Conjugative elements are capable of self-transfer and also capable of mobilizing other DNA elements that are not able to self-transfer. Plasmids that can be mobilized by conjugative elements are generally thought to contain an origin of transfer (oriT), from which mobilization initiates, and to encode a mobilization protein (Mob, a relaxase) that nicks a site in oriT and covalently attaches to the DNA to be transferred. Plasmids that do not have both an oriT and a cognate mob are thought to be nonmobilizable. We found that Bacillus subtilis carrying the integrative and conjugative element ICEBs1 can transfer three different plasmids to recipient bacteria at high frequencies. Strikingly, these plasmids do not have dedicated mobilization-oriT functions. Plasmid mobilization required conjugation proteins of ICEBs1, including the putative coupling protein. In contrast, plasmid mobilization did not require the ICEBs1 conjugative relaxase or cotransfer of ICEBs1, indicating that the putative coupling protein likely interacts with the plasmid replicative relaxase and directly targets the plasmid DNA to the ICEBs1 conjugation apparatus. These results blur the current categorization of mobilizable and nonmobilizable plasmids and indicate that conjugative elements play a role in horizontal gene transfer even more significant than previously recognized.National Institutes of Health (U.S.) (Grant GM50895

A conserved helicase processivity factor is needed for conjugation and replication of an integrative and conjugative element

Integrative and conjugative elements (ICEs) are agents of horizontal gene transfer and have major roles in evolution and acquisition of new traits, including antibiotic resistances. ICEs are found integrated in a host chromosome and can excise and transfer to recipient bacteria via conjugation. Conjugation involves nicking of the ICE origin of transfer (oriT) by the ICE–encoded relaxase and transfer of the nicked single strand of ICE DNA. For ICEBs1 of Bacillus subtilis, nicking of oriT by the ICEBs1 relaxase NicK also initiates rolling circle replication. This autonomous replication of ICEBs1 is critical for stability of the excised element in growing cells. We found a conserved and previously uncharacterized ICE gene that is required for conjugation and replication of ICEBs1. Our results indicate that this gene, helP (formerly ydcP), encodes a helicase processivity factor that enables the host-encoded helicase PcrA to unwind the double-stranded ICEBs1 DNA. HelP was required for both conjugation and replication of ICEBs1, and HelP and NicK were the only ICEBs1 proteins needed for replication from ICEBs1 oriT. Using chromatin immunoprecipitation, we measured association of HelP, NicK, PcrA, and the host-encoded single-strand DNA binding protein Ssb with ICEBs1. We found that NicK was required for association of HelP and PcrA with ICEBs1 DNA. HelP was required for association of PcrA and Ssb with ICEBs1 regions distal, but not proximal, to oriT, indicating that PcrA needs HelP to progress beyond nicked oriT and unwind ICEBs1. In vitro, HelP directly stimulated the helicase activity of the PcrA homologue UvrD. Our findings demonstrate that HelP is a helicase processivity factor needed for efficient unwinding of ICEBs1 for conjugation and replication. Homologues of HelP and PcrA-type helicases are encoded on many known and putative ICEs. We propose that these factors are essential for ICE conjugation, replication, and genetic stability.National Institutes of Health (U.S.) (Grant GM50895

Understanding Pacific Ocean influence on interannual precipitation variability in the Sahel

Moisture budget decomposition is performed for the Sahel (10°–20°N and 20°W–40°E) in order to understand the processes that govern regional hydroclimate variability on interannual time scales and frame them in the context of their primary ocean driver. Results show that warm conditions in the Eastern Tropical Pacific remotely force anomalously dry conditions primarily through affecting the low-troposphere mass divergence field. This behavior is related to increased subsidence over the tropical Atlantic and into the Sahel and an anomalous westward flow of moisture from the continent, both resulting in a coherent drying pattern. Understanding the physical processes relating remote sea surface temperature anomalies to atmospheric circulation changes and the resulting complex local convergence patterns is important for advancing seasonal prediction of precipitation over West Africa

Polar positioning of a conjugation protein from the integrative and conjugative element ICEBs1 of Bacillus subtilis

ICEBs1 is an integrative and conjugative element found in the chromosome of Bacillus subtilis. ICEBs1 encodes functions needed for its excision and transfer to recipient cells. We found that the ICEBs1 gene conE (formerly yddE) is required for conjugation and that conjugative transfer of ICEBs1 requires a conserved ATPase motif of ConE. ConE belongs to the HerA/FtsK superfamily of ATPases, which includes the well-characterized proteins FtsK, SpoIIIE, VirB4, and VirD4. We found that a ConE-GFP (green fluorescent protein) fusion associated with the membrane predominantly at the cell poles in ICEBs1 donor cells. At least one ICEBs1 product likely interacts with ConE to target it to the membrane and cell poles, as ConE-GFP was dispersed throughout the cytoplasm in a strain lacking ICEBs1. We also visualized the subcellular location of ICEBs1. When integrated in the chromosome, ICEBs1 was located near midcell along the length of the cell, a position characteristic of that chromosomal region. Following excision, ICEBs1 was more frequently found near a cell pole. Excision of ICEBs1 also caused altered positioning of at least one component of the replisome. Taken together, our findings indicate that ConE is a critical component of the ICEBs1 conjugation machinery, that conjugative transfer of ICEBs1 from B. subtilis likely initiates at a donor cell pole, and that ICEBs1 affects the subcellular position of the replisome.Public Health Service (grant GM50895 (A.D.G.))Suffolk University. Department of Chemistry and Biochemistr

Micronesian Migrants in Hawaii: Health Issues and Culturally Appropriate, Community-Based Solutions

More than 20,000 Micronesians have migrated to Guam, Hawaii, or the Commonwealth of the Northern Mariana Islands. Of these migrants, more than 8,000 now live in Hawaii. Factors in their home islands driving the recent emigration include the limited economic resources and struggling health care systems. Education systems in Micronesia are inadequate, and there are few job opportunities. The rates of infectious diseases remain high while at the same time, the epidemiologic transition in health has led to an explosion of non-communicable diseases. In Hawaii, the impact of the Micronesians emigration has been significant with most health and education expenses un-reimbursed. The health care costs alone are substantial as many Micronesians travel to Guam or Hawaii for medical treatment unavailable in their home islands. At the same time, Micronesians have difficulty accessing and navigating the health care system. While governmental, private, and academic programs already provide innovative and communitybased services to the Micronesian population, more work remains to be done. Not only are additional services, tailored to the culture and needs of Micronesian migrants, needed but a keener awareness and understanding of the issues surrounding Hawaii’s migrant population must be promoted among all public health stakeholders to ensure that the priority necessary to successfully address these challenges is recognized

Benefits, Facilitators, Barriers, and Strategies to Improve Pesticide Protective Behaviors: Insights from Farmworkers in North Carolina Tobacco Fields

Pesticide exposure is associated with deleterious health effects. Prior studies suggest Latino farmworkers perceive little control over their occupational health. Using the Health Belief Model as a theoretical guide, we explored the perceptions of Latino farmworkers working in tobacco in North Carolina (n = 72) about benefits and facilitators of pesticide protective behaviors as well as barriers, and strategies to overcome barriers to their use. Interviews were conducted with participants at farmworker housing during non-work time. Qualitative data were analyzed using ATLAS.ti. Farmworkers recognized pesticide protective behaviors as helping them to not get sick and stay healthy. Farmworkers perceived work experience as facilitating protective behaviors. Wetness in the field was the most commonly cited barrier to protective behavior use. To overcome this barrier, farmworkers suggested use of water-resistant outerwear, as well as packing a change of clothes for mid-day, with space and time to change provided by employers. Examination of the efficacy and feasibility of farmworkers’ suggestions for addressing barriers is warranted. Training and behavior modeling by experienced peers may improve behavior adoption and perceived control

Invasive Haemophilus influenzae Disease in Adults ≥65 Years, United States, 2011.

BackgroundSince the introduction of the Haemophilus influenzae serotype b vaccine, H influenzae epidemiology has shifted. In the United States, the largest burden of disease is now in adults aged ≥65 years. However, few data exist on risk factors for disease severity and outcome in this age group.MethodsA retrospective case-series review of invasive H influenzae infections in patients aged ≥65 years was conducted for hospitalized cases reported to Active Bacterial Core surveillance in 2011.ResultsThere were 299 hospitalized cases included in the analysis. The majority of cases were caused by nontypeable H influenzae, and the overall case fatality ratio (CFR) was 19.5%. Three or more underlying conditions were present in 63% of cases; 94% of cases had at least 1. Patients with chronic heart conditions (congestive heart failure, coronary artery disease, and/or atrial fibrillation) (odds ratio [OR], 3.27; 95% confidence interval [CI], 1.65-6.46), patients from private residences (OR, 8.75; 95% CI, 2.13-35.95), and patients who were not resuscitate status (OR, 2.72; 95% CI, 1.31-5.66) were more likely to be admitted to the intensive care unit (ICU). Intensive care unit admission (OR, 3.75; 95% CI, 1.71-8.22) and do not resuscitate status (OR, 12.94; 95% CI, 4.84-34.55) were significantly associated with death.ConclusionsWithin this age group, burden of disease and CFR both increased significantly as age increased. Using ICU admission as a proxy for disease severity, our findings suggest several conditions increased risk of disease severity and patients with severe disease were more likely to die. Further research is needed to determine the most effective approach to prevent H influenzae disease and mortality in older adults

Death Notices-Joerg Haeberli, Laura Laurencich Minelli, Ursula Wagner, Nancy Ellen Kirkhuff Porter, Billie Jean Isbell, Lynn Ann Meisch, & Bernard While Bell Jr.

This consists of short biographies of deceased scholars Joerg Haeberli, Laura Laurencich Minelli, Ursula Wagner, Nancy Ellen Kirkhuff Porter, Billie Jean Isbell, Lynn Ann Meisch, and Bernard White Bell Jr

The Iowa Homemaker vol.23, no.5

Moire, Keith Shillington, page 2 Keeping Up With Today, Margaret Ralston, page 3 Christmas Suggestions, Pfc. Schwanz, AS Greenburg, page 4 Navy Menus on Review, Eileen Cooper, page 6 Creating the Holiday Spirit, Joan Miller, page 7 For Christmas I’d Like, Betty Aldrich, page 8 For Use – Peanut Substitutes, Norma Dale, page 10 Wartime Cards Marked by Individuality, Julie Johnston, page 11 What’s New in Home Economics, Lily Houseman, page 12 Iowa Staters Go Caroling, Frances Kerekes, page 16 For the Christmas Stocking¸ Lois Stewart, page 17 European Rationing, Catherine Tidemanson, page 18 Supervise Army Hospital Diets, Virginia Brainard, page 19 Originality Expressed in Festive Wrappings, Doris Gregg, page 20 Across Alumnae Desks, Virginia Carter, page 22 Alums in the News, Rachel Ann Lusher, page 2