Under the stress of ultraviolet radiation some cyanobacteria synthesize scytonemin, a protective pigment against DNA photodamage. In addition to photoprotection, scytonemin has been shown to have an anti-proliferative effect on various types of malignant cells. In this study the effect of scytonemin on melanoma and spleen cells was assessed both in vitro using tissue cultures and in vivo in mice models. Melanoma and spleen cells were exposed to 0.08 to 10 μM of scytonemin, and cell proliferation was measured using tritiated thymidine uptake. The data suggest that scytonemin acts as an inhibitor for melanoma cells in a concentration-dependent manner while enhancing the proliferation of spleen cells, suggesting that it can potentially augment the immune response. Furthermore, mice injected with melanoma cells and scytonemin produced fewer tumors than mice that did not receive scytonemin, although the data were not significant. This study adds to the growing body of research that scytonemin may be beneficial as a future anticancer agent to prevent tumor cell growth.
{"title":"Anti-proliferation of Melanoma Cells and Immune Stimulation by the Cyanobacterial Indole-alkaloid Scytonemin","authors":"A. Jones","doi":"10.33043/ff.7.1.54-63","DOIUrl":"https://doi.org/10.33043/ff.7.1.54-63","url":null,"abstract":"Under the stress of ultraviolet radiation some cyanobacteria synthesize scytonemin, a protective pigment against DNA photodamage. In addition to photoprotection, scytonemin has been shown to have an anti-proliferative effect on various types of malignant cells. In this study the effect of scytonemin on melanoma and spleen cells was assessed both in vitro using tissue cultures and in vivo in mice models. Melanoma and spleen cells were exposed to 0.08 to 10 μM of scytonemin, and cell proliferation was measured using tritiated thymidine uptake. The data suggest that scytonemin acts as an inhibitor for melanoma cells in a concentration-dependent manner while enhancing the proliferation of spleen cells, suggesting that it can potentially augment the immune response. Furthermore, mice injected with melanoma cells and scytonemin produced fewer tumors than mice that did not receive scytonemin, although the data were not significant. This study adds to the growing body of research that scytonemin may be beneficial as a future anticancer agent to prevent tumor cell growth.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82115745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Breanna Brenneman, K. Adamson, M. Beer, Yen-Ning Ho, Kiev S. Gracias, Chelsea M. Priest, Erika N. Biernbaum, J. McKillip
Bacillus cereus is traditionally thought to be the only member of its genus accepted as a pathogen in foods like grains, fruits, vegetables, and milk due to the presence of the nonhemolytic (Nhe) operon. However, many other Bacillus spp. may also harbor the Nhe operon and be pathogenic, including not just food-associated gastrointestinal toxicoinfections, but human endophthalmitis as well. Real-time PCR targeted the nheA gene in 37 samples obtained from food, soil, and reference cultures by analyzing the standard deviations of melt peaks. Repetitive element PCR was used to compare the banding patterns of each sample against B. cereus ATCC 14579 and three B. thuringiensis strains to “fingerprint” each isolate. Of the original 43 isolated tested, 37 were Gram-positive rods. The remaining six samples were Gram-positive cocci. Twenty-five of the 37 Gram-positive Bacillus spp. were nheA positive, while twelve were negative. Many of the nheA positive strains were species not previously known to contain Nhe and were capable of causing gastroenteritis in consumers.
{"title":"Real-time Screening of Foods Using Repetitive Element PCR Reveals a DNA Marker Characteristic for Enterotoxigenic Bacillus Species","authors":"Breanna Brenneman, K. Adamson, M. Beer, Yen-Ning Ho, Kiev S. Gracias, Chelsea M. Priest, Erika N. Biernbaum, J. McKillip","doi":"10.33043/ff.7.1.36-53","DOIUrl":"https://doi.org/10.33043/ff.7.1.36-53","url":null,"abstract":"Bacillus cereus is traditionally thought to be the only member of its genus accepted as a pathogen in foods like grains, fruits, vegetables, and milk due to the presence of the nonhemolytic (Nhe) operon. However, many other Bacillus spp. may also harbor the Nhe operon and be pathogenic, including not just food-associated gastrointestinal toxicoinfections, but human endophthalmitis as well. Real-time PCR targeted the nheA gene in 37 samples obtained from food, soil, and reference cultures by analyzing the standard deviations of melt peaks. Repetitive element PCR was used to compare the banding patterns of each sample against B. cereus ATCC 14579 and three B. thuringiensis strains to “fingerprint” each isolate. Of the original 43 isolated tested, 37 were Gram-positive rods. The remaining six samples were Gram-positive cocci. Twenty-five of the 37 Gram-positive Bacillus spp. were nheA positive, while twelve were negative. Many of the nheA positive strains were species not previously known to contain Nhe and were capable of causing gastroenteritis in consumers.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86115316","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
James C. Kuldell, Harshani Luknauth, Anthony E. Ricigliano, Nathan W. Rigel
The outer membrane is the defining characteristic of Gram-negative bacteria and is crucial for the maintenance of cellular integrity. Lipoproteins are an essential component of this outer membrane and regulate broad cellular functions ranging from efflux, cellular physiology, antibiotic resistance, and pathogenicity. In the canonical model of lipoprotein biogenesis, lipoprotein precursors are first synthesized in the cytoplasm prior to extensive modifications by the consecutive action of three key enzymes: diacylglyceryl transferase (Lgt), lipoprotein signal peptidase A (LspA), and apolipoprotein N-acyltransferase (Lnt). This enzymatic process modifies lipoprotein precursors for subsequent trafficking by the Lol pathway. The function of these three enzymes were originally thought to be essential, however, in some Gram-negative bacteria, namely Acinetobacter baylyi, the third enzyme Lnt is dispensable. Here we review the function and significance of Lgt, LspA, and Lnt in outer membrane biogenesis and how non-canonical models of lipoprotein processing in Acinetobacter spp. can enhance our understanding of lipoprotein modifications and trafficking.
{"title":"Biogenesis of Lipoproteins in Gram-Negative Bacteria: 50 Years of Progress","authors":"James C. Kuldell, Harshani Luknauth, Anthony E. Ricigliano, Nathan W. Rigel","doi":"10.33043/ff.7.1.9-24","DOIUrl":"https://doi.org/10.33043/ff.7.1.9-24","url":null,"abstract":"The outer membrane is the defining characteristic of Gram-negative bacteria and is crucial for the maintenance of cellular integrity. Lipoproteins are an essential component of this outer membrane and regulate broad cellular functions ranging from efflux, cellular physiology, antibiotic resistance, and pathogenicity. In the canonical model of lipoprotein biogenesis, lipoprotein precursors are first synthesized in the cytoplasm prior to extensive modifications by the consecutive action of three key enzymes: diacylglyceryl transferase (Lgt), lipoprotein signal peptidase A (LspA), and apolipoprotein N-acyltransferase (Lnt). This enzymatic process modifies lipoprotein precursors for subsequent trafficking by the Lol pathway. The function of these three enzymes were originally thought to be essential, however, in some Gram-negative bacteria, namely Acinetobacter baylyi, the third enzyme Lnt is dispensable. Here we review the function and significance of Lgt, LspA, and Lnt in outer membrane biogenesis and how non-canonical models of lipoprotein processing in Acinetobacter spp. can enhance our understanding of lipoprotein modifications and trafficking.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88277804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Competitive runners experience various risk factors that render them more susceptible to superficial cutaneous fungal infections, including the use of occlusive footwear, shared locker rooms, submission of feet to constant maceration, trauma, sweating, and having depressed immune function. The goal of this work was to assess the prevalence of athlete’s foot fungi in cross country runners at St. John Fisher College. Toe webs of 16 collegiate runners were sampled and volunteers surveyed about their shoe habits, foot hygiene, and average miles run per week. Lack of tinea pedis-causing fungi in asymptomatic cross- country runners shifted the study to investigate the identities of fungi morphologically similar to athlete’s foot and look for correlations with volunteers’ running habits and hygiene. Thirty-five distinct fungal cultures were isolated and compared to a known Trichophyton rubrum strain both microscopically and macroscopically. Four samples were preliminarily identified as tinea pedis-causing fungi and sequenced to confirm molecular identification. Fungal DNA was isolated, purified, and PCR amplified using primers for the internal transcribed spacer region, D1/D2 region of the 28S subunit, and β-Tubulin gene. Three of the four isolates were identified as Fusarium equiseti, a soil-borne plant pathogen with rare human pathogenicity reported. The fourth isolate was Beauveria bassiana, a common soil-borne pathogen that can infect immunocompromised individuals. Correct dermatophytic identification and understanding of the interplay between species is important to provide correct treatment, prevent spread among athletes and within facilities, and determine how opportunistic pathogens might play a role in people with immune suppressed function, which includes runners.
{"title":"Isolation and Identification of Dermatophytes from Collegiate Runners","authors":"L. Kalnina, Stephanie Guzelak, Maryann Herman","doi":"10.33043/ff.7.1.64-73","DOIUrl":"https://doi.org/10.33043/ff.7.1.64-73","url":null,"abstract":"Competitive runners experience various risk factors that render them more susceptible to superficial cutaneous fungal infections, including the use of occlusive footwear, shared locker rooms, submission of feet to constant maceration, trauma, sweating, and having depressed immune function. The goal of this work was to assess the prevalence of athlete’s foot fungi in cross country runners at St. John Fisher College. Toe webs of 16 collegiate runners were sampled and volunteers surveyed about their shoe habits, foot hygiene, and average miles run per week. Lack of tinea pedis-causing fungi in asymptomatic cross- country runners shifted the study to investigate the identities of fungi morphologically similar to athlete’s foot and look for correlations with volunteers’ running habits and hygiene. Thirty-five distinct fungal cultures were isolated and compared to a known Trichophyton rubrum strain both microscopically and macroscopically. Four samples were preliminarily identified as tinea pedis-causing fungi and sequenced to confirm molecular identification. Fungal DNA was isolated, purified, and PCR amplified using primers for the internal transcribed spacer region, D1/D2 region of the 28S subunit, and β-Tubulin gene. Three of the four isolates were identified as Fusarium equiseti, a soil-borne plant pathogen with rare human pathogenicity reported. The fourth isolate was Beauveria bassiana, a common soil-borne pathogen that can infect immunocompromised individuals. Correct dermatophytic identification and understanding of the interplay between species is important to provide correct treatment, prevent spread among athletes and within facilities, and determine how opportunistic pathogens might play a role in people with immune suppressed function, which includes runners.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86447118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Avery M Runnebohm, Melissa D Evans, Adam E Richardson, Samantha M Turk, James B Olesen, Philip J Smaldino, Eric M Rubenstein
Ubr1 is a conserved ubiquitin ligase involved in the degradation of aberrant proteins in eukaryotic cells. The human enzyme is found mutated in patients with Johanson-Blizzard syndrome. We hypothesized that Ubr1 is necessary for optimal cellular fitness in conditions associated with elevated abundance of aberrant and misfolded proteins. Indeed, we found that loss of Ubr1 in the model eukaryotic microorganism Saccharomyces cerevisiae strongly sensitizes cells to hygromycin B, which reduces translational fidelity by causing ribosome A site distortion. Our results are consistent with a prominent role for Ubr1 in protein quality control. We speculate that disease manifestations in patients with Johanson-Blizzard syndrome are linked, at least in part, to defects in protein quality control caused by loss of Ubr1 function.
{"title":"Loss of protein quality control gene <i>UBR1</i> sensitizes <i>Saccharomyces cerevisiae</i> to the aminoglycoside hygromycin B.","authors":"Avery M Runnebohm, Melissa D Evans, Adam E Richardson, Samantha M Turk, James B Olesen, Philip J Smaldino, Eric M Rubenstein","doi":"10.33043/FF.6.1.76-83","DOIUrl":"https://doi.org/10.33043/FF.6.1.76-83","url":null,"abstract":"<p><p>Ubr1 is a conserved ubiquitin ligase involved in the degradation of aberrant proteins in eukaryotic cells. The human enzyme is found mutated in patients with Johanson-Blizzard syndrome. We hypothesized that Ubr1 is necessary for optimal cellular fitness in conditions associated with elevated abundance of aberrant and misfolded proteins. Indeed, we found that loss of Ubr1 in the model eukaryotic microorganism <b><i>Saccharomyces cerevisiae</i></b> strongly sensitizes cells to hygromycin B, which reduces translational fidelity by causing ribosome A site distortion. Our results are consistent with a prominent role for Ubr1 in protein quality control. We speculate that disease manifestations in patients with Johanson-Blizzard syndrome are linked, at least in part, to defects in protein quality control caused by loss of Ubr1 function.</p>","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7861132/pdf/nihms-1648971.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25343303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The benefits of mentoring undergraduate research students","authors":"Douglas H. Roossien","doi":"10.33043/FF.6.1.9-11","DOIUrl":"https://doi.org/10.33043/FF.6.1.9-11","url":null,"abstract":"Faculty Perspective by Dr. Douglas H. Roossien","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78520175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
John A. Haradean, Tyler Ralph-Epps, Zach Whiteacre, S. Neumann, D. Becker
Bioremediation is currently under investigation as a viable way to remove many environmental pollutants and most commonly involves the use of microorganisms to extract organic pollutants or heavy metals from water or soil. One of the most abundant heavy metals found in industrially polluted sites is zinc (Zn); it is often found alongside metals like lead (Pb), arsenic (As), and mercury (Hg). This experiment investigated the potential bioremediation of pasteurized soil contaminated with zinc using different vesicular arbuscular mycorrhizal fungi (VAM) species and lettuce plants (Lactuca sativa). Soil was amended with 0.4 g of zinc chloride (ZnCl2) per kg of soil. Amended and unamended soils were inoculated with two different mixes of VAM, BioAg VAM-Endo™ and MycoBloom. For each treatment, L. sativa plants (15 pots per treatment) were grown in a greenhouse setting. Plant diameter was measured weekly. Plants were harvested after 55-days and the wet weight of leaf tissue was measured before the tissue was sent for analysis of zinc levels. Roots were assessed for mycorrhizae using a trypan blue staining procedure. The BioAg VAM-Endo™ mix was the most successful at removing ZnCl2 from the soil. L. sativa inoculated with VAM mixes formed mycorrhizae, grew healthier and removed more zinc from the soil than the non-inoculated group. We propose further investigation into the use of mycorrhizal fungi paired with other plant species to remove zinc from contaminated sites with harmful levels of zinc.
{"title":"Mixtures of Mycorrhizal Fungi Improve Growth of Lactuca Sativa and Reduce Levels of Zinc in Contaminated Soil","authors":"John A. Haradean, Tyler Ralph-Epps, Zach Whiteacre, S. Neumann, D. Becker","doi":"10.33043/ff.5.1.65-74","DOIUrl":"https://doi.org/10.33043/ff.5.1.65-74","url":null,"abstract":"Bioremediation is currently under investigation as a viable way to remove many environmental pollutants and most commonly involves the use of microorganisms to extract organic pollutants or heavy metals from water or soil. One of the most abundant heavy metals found in industrially polluted sites is zinc (Zn); it is often found alongside metals like lead (Pb), arsenic (As), and mercury (Hg). This experiment investigated the potential bioremediation of pasteurized soil contaminated with zinc using different vesicular arbuscular mycorrhizal fungi (VAM) species and lettuce plants (Lactuca sativa). Soil was amended with 0.4 g of zinc chloride (ZnCl2) per kg of soil. Amended and unamended soils were inoculated with two different mixes of VAM, BioAg VAM-Endo™ and MycoBloom. For each treatment, L. sativa plants (15 pots per treatment) were grown in a greenhouse setting. Plant diameter was measured weekly. Plants were harvested after 55-days and the wet weight of leaf tissue was measured before the tissue was sent for analysis of zinc levels. Roots were assessed for mycorrhizae using a trypan blue staining procedure. The BioAg VAM-Endo™ mix was the most successful at removing ZnCl2 from the soil. L. sativa inoculated with VAM mixes formed mycorrhizae, grew healthier and removed more zinc from the soil than the non-inoculated group. We propose further investigation into the use of mycorrhizal fungi paired with other plant species to remove zinc from contaminated sites with harmful levels of zinc.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81313366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
While healthcare professionals are working in hospitals, they will often manipulate the privacy curtains during the care of their patients. Studies have shown that the transfer of bacteria from hands to the curtains and vice versa is possible. Despite the possibility of hospital curtains being a mode of infection transmission, studies have shown that 53% of hospitals surveyed did not have a policy for cleaning or changing their curtains. The question that this study focused on was whether curtain material affects the persistence of Staphylococcus aureus. In this study, five different curtain types were inoculated with overnight, diluted, and finger imprint cultures of S. aureus. They were incubated at room temperature and were sampled for growth regularly onto Mannitol Salt Agar plates. The colonies were counted, and one-way ANOVA statistical analysis was completed on the data. The statistical analysis showed that the length of persistence of liquid cultures of S. aureus on the curtains was not dependent upon initial concentration. Finger imprint inoculations of four curtain varieties had statistically significant longer persistence times than the liquid cultures. Only the curtain type composed of 100% antimicrobial polyester with water repellant had significantly lower persistence times for the finger imprint culture than the other four curtains. The results suggest that the 100% inherently flame resistant antimicrobial polyester curtain material reduces S. aureus persistence times and that it may benefit hospitals to use this type of curtain.
{"title":"The Persistence of Staphylococcus aureus on Hospital Privacy Curtains","authors":"Sarah E. Cole, Brittany J. Gasper","doi":"10.33043/ff.5.1.53-62","DOIUrl":"https://doi.org/10.33043/ff.5.1.53-62","url":null,"abstract":"While healthcare professionals are working in hospitals, they will often manipulate the privacy curtains during the care of their patients. Studies have shown that the transfer of bacteria from hands to the curtains and vice versa is possible. Despite the possibility of hospital curtains being a mode of infection transmission, studies have shown that 53% of hospitals surveyed did not have a policy for cleaning or changing their curtains. The question that this study focused on was whether curtain material affects the persistence of Staphylococcus aureus. In this study, five different curtain types were inoculated with overnight, diluted, and finger imprint cultures of S. aureus. They were incubated at room temperature and were sampled for growth regularly onto Mannitol Salt Agar plates. The colonies were counted, and one-way ANOVA statistical analysis was completed on the data. The statistical analysis showed that the length of persistence of liquid cultures of S. aureus on the curtains was not dependent upon initial concentration. Finger imprint inoculations of four curtain varieties had statistically significant longer persistence times than the liquid cultures. Only the curtain type composed of 100% antimicrobial polyester with water repellant had significantly lower persistence times for the finger imprint culture than the other four curtains. The results suggest that the 100% inherently flame resistant antimicrobial polyester curtain material reduces S. aureus persistence times and that it may benefit hospitals to use this type of curtain.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76993975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Justin Langro, Megan M. Chamberland, Celena M. Gwin, N. Prakash, D. Velez, Nathan W. Rigel
Protein export pathways are important for bacterial physiology among pathogens and non-pathogens alike. This includes the Twin-Arginine Translocation (Tat) pathway, which transports fully folded proteins across the bacterial cytoplasmic membrane. Some Tat substrates are virulence factors, while others are important for cellular processes like peptidoglycan remodeling. Some bacteria encode more than one copy of each Tat component, including the Gram-negative soil isolate Acinetobacter baylyi. One of these Tat pathways is essential for growth, while the other is not. We constructed a loss-of-function mutation to disrupt the non-essential tatC2 gene and assessed its contribution to cell growth under different environmental conditions. While the tatC2 mutant grew well under standard laboratory conditions, it displayed a growth defect and an aberrant cellular morphology when subjected to high temperature stress including an aberrant cellular morphology. Furthermore, increased sensitivities to detergent suggested a compromised cell envelope. Lastly, using an in vitro co-culture system, we demonstrate that the non-essential Tat pathway provides a growth advantage. The findings of this study establish the importance of the non-essential Tat pathway for optimal growth of A. baylyi in stressful environmental conditions.
{"title":"TatC2 is Important for Growth of Acinetobacter baylyi Under Stress Conditions","authors":"Justin Langro, Megan M. Chamberland, Celena M. Gwin, N. Prakash, D. Velez, Nathan W. Rigel","doi":"10.33043/ff.5.1.37-50","DOIUrl":"https://doi.org/10.33043/ff.5.1.37-50","url":null,"abstract":"Protein export pathways are important for bacterial physiology among pathogens and non-pathogens alike. This includes the Twin-Arginine Translocation (Tat) pathway, which transports fully folded proteins across the bacterial cytoplasmic membrane. Some Tat substrates are virulence factors, while others are important for cellular processes like peptidoglycan remodeling. Some bacteria encode more than one copy of each Tat component, including the Gram-negative soil isolate Acinetobacter baylyi. One of these Tat pathways is essential for growth, while the other is not. We constructed a loss-of-function mutation to disrupt the non-essential tatC2 gene and assessed its contribution to cell growth under different environmental conditions. While the tatC2 mutant grew well under standard laboratory conditions, it displayed a growth defect and an aberrant cellular morphology when subjected to high temperature stress including an aberrant cellular morphology. Furthermore, increased sensitivities to detergent suggested a compromised cell envelope. Lastly, using an in vitro co-culture system, we demonstrate that the non-essential Tat pathway provides a growth advantage. The findings of this study establish the importance of the non-essential Tat pathway for optimal growth of A. baylyi in stressful environmental conditions.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75479068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ethan S. Pickerill, Caleb Embree, Ben A. Evans, E. North, G. M. Mager, D. Bernstein
In 2010, the CRISPR/Cas system of Streptococcus thermophilus was found necessary and sufficient to cleave bacteriophage DNA. Since this time, CRISPR went from a niche scientific field to the laboratories of major research institutions, undergraduate classrooms, and popular culture. In the future, CRISPR may stand along with PCR, DNA sequencing, and transformation as paradigm shifting discoveries in molecular biology. CRISPR genome editing is technically uncomplicated and relatively inexpensive. Thus, CRISPR-mediated genome editing has been adopted by and applied to undergraduate curricula in a wide variety of ways. In this review, we provide an overview of CRISPR-mediated genome editing and examine some of the ways this technology is being leveraged to train students in the classroom and laboratory.
{"title":"How CRISPR-Mediated Genome Editing is Affecting Undergraduate Biology Education","authors":"Ethan S. Pickerill, Caleb Embree, Ben A. Evans, E. North, G. M. Mager, D. Bernstein","doi":"10.33043/ff.5.1.23-34","DOIUrl":"https://doi.org/10.33043/ff.5.1.23-34","url":null,"abstract":"In 2010, the CRISPR/Cas system of Streptococcus thermophilus was found necessary and sufficient to cleave bacteriophage DNA. Since this time, CRISPR went from a niche scientific field to the laboratories of major research institutions, undergraduate classrooms, and popular culture. In the future, CRISPR may stand along with PCR, DNA sequencing, and transformation as paradigm shifting discoveries in molecular biology. CRISPR genome editing is technically uncomplicated and relatively inexpensive. Thus, CRISPR-mediated genome editing has been adopted by and applied to undergraduate curricula in a wide variety of ways. In this review, we provide an overview of CRISPR-mediated genome editing and examine some of the ways this technology is being leveraged to train students in the classroom and laboratory.","PeriodicalId":87255,"journal":{"name":"Fine focus","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90596499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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