Loss of PopZ At activity in Agrobacterium tumefaciens by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects.

Agrobacterium tumefaciens grows by addition of peptidoglycan (PG) at one pole of the bacterium. During the cell cycle, the cell needs to maintain two different developmental programs, one at the growth pole and another at the inert old pole. Proteins involved in this process are not yet well characterized. To further characterize the role of pole-organizing protein A. tumefaciens PopZ (PopZ At ), we created deletions of the five PopZ At domains and assayed their localization. In addition, we created a popZAt deletion strain (ΔpopZAt ) that exhibited growth and cell division defects with ectopic growth poles and minicells, but the strain is unstable. To overcome the genetic instability, we created an inducible PopZ At strain by replacing the native ribosome binding site with a riboswitch. Cultivated in a medium without the inducer theophylline, the cells look like ΔpopZAt cells, with a branching and minicell phenotype. Adding theophylline restores the wild-type (WT) cell shape. Localization experiments in the depleted strain showed that the domain enriched in proline, aspartate, and glutamate likely functions in growth pole targeting. Helical domains H3 and H4 together also mediate polar localization, but only in the presence of the WT protein, suggesting that the H3 and H4 domains multimerize with WT PopZ At , to stabilize growth pole accumulation of PopZ AtIMPORTANCEAgrobacterium tumefaciens is a rod-shaped bacterium that grows by addition of PG at only one pole. The factors involved in maintaining cell asymmetry during the cell cycle with an inert old pole and a growing new pole are not well understood. Here we investigate the role of PopZ At , a homologue of Caulobacter crescentus PopZ (PopZ Cc ), a protein essential in many aspects of pole identity in C. crescentus We report that the loss of PopZ At leads to the appearance of branching cells, minicells, and overall growth defects. As many plant and animal pathogens also employ polar growth, understanding this process in A. tumefaciens may lead to the development of new strategies to prevent the proliferation of these pathogens. In addition, studies of A. tumefaciens will provide new insights into the evolution of the genetic networks that regulate bacterial polar growth and cell division

Patterns Of Human Papillomavirus (hpv) Infection In Female Teenagers In Buri Ram And Udon Thani, Thailand: A Secondary Data Analysis

Background Cervical cancer is the second most common cancer with an age-standardized incidence rate (ASR) of 16.2/100000 person-year among Thailand women. Human papillomavirus (HPV) is a necessary cause of cervical cancer, and HPV-related cancers have a substantial global burden accounting for approximately 5.2% of all cancers worldwide. Since the prevalence of HPV genotypes can be different by areas, it is critical to understand regional- and population-level HPV prevalence before establishing vaccination programs. Methods Data were obtained in August 2019 from the effectiveness study of a one vs. two doses of bivalent HPV vaccine in female teenagers in two provinces in Thailand. Female students were recruited from regular high schools for Grade 10 and Grade 12 (age 16-18 years) and vocational schools for Year 1 and Year 3 (age 16-18 years) corresponding to Grade 10 and 12 in each province. A urine sample was collected at school for HPV DNA PCR genotyping from all participants. HPV infection was determined in urine samples by PCR DNA assay and Cobas 4800 HPV test. The geospatial visualization was performed to compare the prevalence of overall HPV and HPV genotypes between districts in two provinces. To estimate differences in the prevalence of HPV infection and genotypes between the cohorts in each category (i.e. grade and school type), the Analysis of Variance (ANOVA) test was performed. Results A total of 8,594 female students were recruited in the study. 4,205 were from Buri Ram, and 4,389 were from Udon Thani. In total, the overall HPV prevalence was 12.5%. The prevalence of HPV16, HPV18, and other HPV genotypes was 3.5%, 2.0%, and 7.6%, respectively. In Buri Ram, Chalermphrakiet district had the highest HPV prevalence (23.1%), and Cham Ni district had the lowest HPV prevalence (0.0%) out of 23 districts. In Udon Thani, Ban Dung district had the highest HPV prevalence (16.9%), and K. Ku Kaeo district had the lowest HPV prevalence (9.0%) out of 20 districts. The prevalence of overall HPV and HPV genotypes increased with grade and was higher in vocational school compared to regular school. Conclusions In conclusion, this secondary data analysis reports the prevalence of HPV genotypes in female teenagers in Buri Ram and Udon Thani provinces, Thailand. In both provinces, HPV prevalence was significantly different between grades and school types regardless of genotypes. HPV prevalence increased with grade and the vocational school had higher HPV prevalence than regular school type

Loss of PopZ At activity in Agrobacterium tumefaciens by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects.

Agrobacterium tumefaciens grows by addition of peptidoglycan (PG) at one pole of the bacterium. During the cell cycle, the cell needs to maintain two different developmental programs, one at the growth pole and another at the inert old pole. Proteins involved in this process are not yet well characterized. To further characterize the role of pole-organizing protein A. tumefaciens PopZ (PopZ At ), we created deletions of the five PopZ At domains and assayed their localization. In addition, we created a popZAt deletion strain (ΔpopZAt ) that exhibited growth and cell division defects with ectopic growth poles and minicells, but the strain is unstable. To overcome the genetic instability, we created an inducible PopZ At strain by replacing the native ribosome binding site with a riboswitch. Cultivated in a medium without the inducer theophylline, the cells look like ΔpopZAt cells, with a branching and minicell phenotype. Adding theophylline restores the wild-type (WT) cell shape. Localization experiments in the depleted strain showed that the domain enriched in proline, aspartate, and glutamate likely functions in growth pole targeting. Helical domains H3 and H4 together also mediate polar localization, but only in the presence of the WT protein, suggesting that the H3 and H4 domains multimerize with WT PopZ At , to stabilize growth pole accumulation of PopZ AtIMPORTANCEAgrobacterium tumefaciens is a rod-shaped bacterium that grows by addition of PG at only one pole. The factors involved in maintaining cell asymmetry during the cell cycle with an inert old pole and a growing new pole are not well understood. Here we investigate the role of PopZ At , a homologue of Caulobacter crescentus PopZ (PopZ Cc ), a protein essential in many aspects of pole identity in C. crescentus We report that the loss of PopZ At leads to the appearance of branching cells, minicells, and overall growth defects. As many plant and animal pathogens also employ polar growth, understanding this process in A. tumefaciens may lead to the development of new strategies to prevent the proliferation of these pathogens. In addition, studies of A. tumefaciens will provide new insights into the evolution of the genetic networks that regulate bacterial polar growth and cell division