ABSTRACT
Tn7-like transposons are characterized by their ability to insert specifically into host chromosomes. Recognition of the attachment (att) site by TnsD recruits the TnsABC proteins to form the transpososome and facilitate transposition. Although this pathway is well established, atomic-level structural insights of this process remain largely elusive. Here, we present the cryo-electron microscopy (cryo-EM) structures of the TnsC-TnsD-att DNA complex and the TnsABCD transpososome from the Tn7-like transposon in Peltigera membranacea cyanobiont 210A, a type I-B CRISPR-associated transposon. Our structures reveal a striking bending of the att DNA, featured by the intercalation of an arginine side chain of TnsD into a CC/GG dinucleotide step. The TnsABCD transpososome structure reveals TnsA-TnsB interactions and demonstrates that TnsC not only recruits TnsAB but also directly participates in the transpososome assembly. These findings provide mechanistic insights into targeted DNA insertion by Tn7-like transposons, with implications for improving the precision and efficiency of their genome-editing applications.
ABSTRACT
Tn7-like transposons have co-opted CRISPR-Cas systems to facilitate the movement of their own DNA. These CRISPR-associated transposons (CASTs) are promising tools for programmable gene knockin. A key feature of CASTs is their ability to recruit Tn7-like transposons to nuclease-deficient CRISPR effectors. However, how Tn7-like transposons are recruited by diverse CRISPR effectors remains poorly understood. Here, we present the cryo-EM structure of a recruitment complex comprising the Cascade complex, TniQ, TnsC, and the target DNA in the type I-B CAST from Peltigera membranacea cyanobiont 210A. Target DNA recognition by Cascade induces conformational changes in Cas6 and primes TniQ recruitment through its C-terminal domain. The N-terminal domain of TniQ is bound to the seam region of the TnsC spiral heptamer. Our findings provide insights into the diverse mechanisms for the recruitment of Tn7-like transposons to CRISPR effectors and will aid in the development of CASTs as gene knockin tools.
Subject(s)
Ascomycota , CRISPR-Associated Proteins , CRISPR-Cas Systems , DNA Transposable Elements , Gene Knock-In Techniques , CRISPR-Associated Proteins/metabolism , CRISPR-Associated Proteins/ultrastructure , Cryoelectron Microscopy , Ascomycota/chemistry , Ascomycota/metabolism , Ascomycota/ultrastructureABSTRACT
ResumenEl objetivo de este artículo es investigar el posible papel desempeñado por los teléfonos móviles como depósitos de colonización bacteriana y los factores de riesgo que ésta conlleva en un ambiente hospitalario. Entre enero de 2013 y marzo de 2014 examinamos a 226 miembros del personal de un hospital regional de Australia (146 médicos y 80 estudiantes de medicina). Los principales resultados de interés se relacionaron con los tipos de microorganismos y la cantidad de contaminación encontrados en los teléfonos móviles. Este estudio mostró la existencia de un alto nivel de contaminación bacteriana (n = 168/226, 74%) en los teléfonos móviles de los funcionarios de un hospital de atención terciaria, aislándose organismos similares en la mano dominante del personal y en sus teléfonos móviles. Mientras que la mayoría de los organismos aislados pertenecía a la flora cutánea normal, un pequeño porcentaje era potencialmente patógeno (n = 12/226, 5%). Además, se encontró que ser miembro subalterno del personal médico constituía un factor de riesgo para un importante crecimiento microbiano (OR 4.00, 95% CI 1.54, 10.37). Sólo 31% (70/226) de los participantes en el estudio informó que limpiaba sus teléfonos regularmente y sólo 21% (47/226) reportó que usa toallitas con alcohol para la limpieza de su teléfono. Este estudio demuestra que los teléfonos móviles son potenciales vehículos de bacterias patógenas en un ambiente hospitalario. Sólo una minoría de participantes informó que limpia su teléfono regularmente. Deberían elaborarse y aplicarse directrices de desinfección utilizando toallitas con alcohol.
Subject(s)
Retrospective Studies , AustraliaABSTRACT
The objective of this article is to investigate the potential role of mobile phones as a reservoir for bacterial colonization and the risk factors for bacterial colonization in a hospital setting. We screened 226 staff members at a regional Australian hospital (146 doctors and 80 medical students) between January 2013 and March 2014. The main outcomes of interest were the types of microorganisms and the amount of contamination of the mobile phones. This study found a high level of bacterial contamination (n = 168/226, 74%) on the mobile phones of staff members in a tertiary hospital, with similar organisms isolated from the staff member's dominant hand and mobile phones. While most of the isolated organisms were normal skin flora, a small percentage were potentially pathogenic (n = 12/226, 5%). Being a junior medical staff was found to be a risk factor for heavy microbial growth (OR 4.00, 95% CI 1.54, 10.37). Only 31% (70/226) of our participants reported cleaning their phones routinely, and only 21% (47/226) reported using alcohol containing wipes on their phones. This study demonstrates that mobile phones are potentially vehicles for pathogenic bacteria in a hospital setting. Only a minority of our participants reported cleaning their phones routinely. Disinfection guidelines utilizing alcohol wipes should be developed and implemented.
Subject(s)
Alcohols , Bacteria/isolation & purification , Bacterial Infections/prevention & control , Cell Phone , Cross Infection/prevention & control , Disinfectants , Fomites/microbiology , Hand/microbiology , Australia , Bacteria/classification , Bacterial Infections/microbiology , Bacterial Infections/transmission , Colony Count, Microbial , Cross Infection/microbiology , Cross Infection/transmission , Drug Delivery Systems , Female , Humans , Male , Medical Staff, Hospital , Risk Factors , Students, Medical , Tertiary Care CentersABSTRACT
Due to its overall environmental impact, the residual dye in the wastewater from the synthetic dye manufacturing and textile industries is a global concern. The discharge contains a high content of pigments and other additives, possessing complex structures. As per the requirement for dyed clothing, dyestuff in the effluent is less susceptible to acids, bases, and oxygen. Thus, conventional physical and chemical methods are not always efficient in degrading the dyes. Some microorganisms growing in an area affected with textile effluent have the capability to utilize the dyes as a source of carbon or nitrogen or both. As a very clean, inexpensive, and sufficient alternative, bioremediation of textile wastewater using these microorganisms has gained major popularity. This review primarily centers the contribution of bacteria in this sector and the isolation of such bacteria from textile effluent. A secondary focus is discussing the factors which influence the performance by different bacteria.
ABSTRACT
Urinary Tract Infection (UTI) is a worldwide phenomenon in modern times, in which the dependency on antibiotics for its treatment is increasing. The current study was conducted in order to find alternatives to antibiotics by investigating some commercial fruits for their antimicrobial activity. The fruits in this study included green apple (Malus domestica), papaya (Carica papaya), lemon (Citrus limon), and strawberry (Fragaria ananassa), which were used to prepare methanolic and ethanolic extracts through Soxhlet extraction technique. The extracts were used against bacteria that cause UTI, and five different strains were selected: E. coli (ATCC: 15922), E. coli (ATCC: 25922), Pseudomonas aeruginosa (ATCC: 27853), Enterococcus faecalis (ATCC: 29212), and Klebsiella pneumoniae. Antimicrobial tests of the extracts were conducted by following the agar well diffusion method, where ciprofloxacin was used as a positive control, and autoclaved distilled water was used as a negative control. Among the fruits, apple and papaya extracts did not show any zone of inhibition against any of the tested bacteria. However, both lemon and strawberry extracts showed inhibition zone against all of the mentioned bacteria. The ethanolic extracts of lemon and strawberry were more potent than their methanolic extracts. Lemon ethanolic extract showed the highest zone of inhibition against Pseudomonas aeruginosa (ATCC: 27853) (18.34 ± 0.58) and lowest one against Klebsiella pneumoniae (16.00 ± 1.00). Strawberry ethanolic extracts showed the highest zone of inhibition against Pseudomonas aeruginosa (ATCC: 27853) (16.33 ± 0.58) and the lowest one against Klebsiella pneumoniae (13.33 ± 0.58). As antibiotic resistance is paving the way for multi-drug resistant bacteria, the results of lemon and strawberry can be considered to be used as an antimicrobial agent in treating urinary tract infections.
ABSTRACT
OBJECTIVE: Halotolerant bacteria have multiple uses viz. fermentation with lesser sterility control and industrial production of bioplastics. Moreover, it may increase the crop productivity of coastal saline lands in Bangladesh by transferring the salt tolerant genes into the plants. The study focused on the isolation and identification of the halotolerant bacteria from three soil samples, collected from coastal Patenga area. The samples were inoculated in nutrient media containing a wide range of salt concentrations. RESULTS: All the samples showed 2, 4 and 6% (w/v) salt tolerance. The isolates from Patenga soil (4, 6%) and beach soil (2%) showed catalase activity and all the isolates showed negative results for oxidase activity, indole production, lactose and motility. All the samples provided positive results for dextrose fermentation. Other tests provided mixed results. Based on the morphological characteristics, biochemical tests and ABIS software analysis the isolates fall within the Enterobacteriaceae, Clostridium and Corynebacterium, with a predominance of Vibrios. Overall the isolates can be considered as mild halotolerant, with the best growth observed at lower salinities and no halophilism detected. Among many possibilities, the genes responsible for the salt tolerant trait in these species can be identified, extracted and inserted into the crop plants to form a transgenic plant to result in higher yield for the rest of the year.