Reconstitution of a minimal DNA replicase from Pseudomonas aeruginosa and stimulation by non-cognate auxiliary factors.

نویسندگان

  • Thale C Jarvis
  • Amber A Beaudry
  • James M Bullard
  • Nebojsa Janjic
  • Charles S McHenry
چکیده

DNA polymerase III holoenzyme is responsible for chromosomal replication in bacteria. The components and functions of Escherichia coli DNA polymerase III holoenzyme have been studied extensively. Here, we report the reconstitution of replicase activity by essential components of DNA polymerase holoenzyme from the pathogen Pseudomonas aeruginosa. We have expressed and purified the processivity factor (beta), single-stranded DNA-binding protein, a complex containing the polymerase (alpha) and exonuclease (epsilon) subunits, and the essential components of the DnaX complex (tau(3)deltadelta'). Efficient primer elongation requires the presence of alphaepsilon, beta, and tau(3)deltadelta'. Pseudomonas aeruginosa alphaepsilon can substitute completely for E. coli polymerase III in E. coli holoenzyme reconstitution assays. Pseudomonas beta and tau(3)deltadelta' exhibit a 10-fold lower activity relative to their E. coli counterparts in E. coli holoenzyme reconstitution assays. Although the Pseudomonas counterpart to the E. coli psi subunit was not apparent in sequence similarity searches, addition of purified E. coli chi and psi (components of the DnaX complex) increases the apparent specific activity of the Pseudomonas tau(3)deltadelta' complex approximately 10-fold and enables the reconstituted enzyme to function better under physiological salt conditions.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Investigating the Presence of Type IV Pilin Subgenus in Pseudomonas Aeruginosa Isolated from Clinical and Non-Clinical Samples

Introduction: The type IV Pilin is an important colonization factor for opportunistic pathogens of Pseudomonas aeruginosa, which plays a role in the formation of biofilms and binding to the host cells. Each type of Pilin is coded with a particular auxiliary gene. This specific relationship can be used as a therapeutic target for detecting Pseudomonas aeruginosa strains as well as its molecular ...

متن کامل

Antimicrobial characteristics of some herbal Oils on Pseudomonas aeruginosa with special reference to their chemical compositions

Pseudomonas aeruginosa is an important opportunistic pathogen causing widespread infections by numerous virulence factors. Increasing resistance to antibiotics makes the Pseudomonas infections treatment further difficult. The purpose of this study was to evaluate antimicrobial characteristics of essential oils from Matricaria chamomilla, Artemisia persica, Zataria multiflora, Myrtus communis, R...

متن کامل

Antimicrobial characteristics of some herbal Oils on Pseudomonas aeruginosa with special reference to their chemical compositions

Pseudomonas aeruginosa is an important opportunistic pathogen causing widespread infections by numerous virulence factors. Increasing resistance to antibiotics makes the Pseudomonas infections treatment further difficult. The purpose of this study was to evaluate antimicrobial characteristics of essential oils from Matricaria chamomilla, Artemisia persica, Zataria multiflora, Myrtus communis, R...

متن کامل

Imipenem-resistant Pseudomonas aeruginosa strains carry vim-type metallo-beta-lactamases isolated from intensive care unit, Shahid Beheshti Hospital, North of Iran

Background: Pseudomonas aeruginosa is the causing agent of many hospital infections and metallo-beta-lactamases (MBL) are being reported with increasing frequency. The aim of this study was to determine the frequency of metallo-&beta-lactamases (MBL) and VIM-1 gene in multidrug-resistant strains of P. aeruginosa isolates and to compare the methods of phenotypic and molecular detection. Materia...

متن کامل

A Novel High-Performance Liquid Chromatography Method for Detection of Alginate in Pseudomonas aeruginosa

Background and Objective: The opportunistic pathogen Pseudomonas aeruginosa secrets a capsule-like polysaccharide called alginate which is important for evasion of host defenses, especially in patients with suppressed immunity. Method of alginate determination has an important role in the study of microb...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • The Journal of biological chemistry

دوره 280 9  شماره 

صفحات  -

تاریخ انتشار 2005