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Forkhead-associated proteins genetically linked to the serine/threonine kinase PknB regulate carbon flux towards antibiotic biosynthesis in Streptomyces coelicolor.

Jones G, Del Sol R, Dudley E, Dyson P - Microb Biotechnol (2010)

Bottom Line: Here we report functional analysis of pknB and two linked genes, fhaAB, encoding forkhead-associated (FHA) domain proteins that are part of a highly conserved gene locus in actinobacteria.FhaAB are candidate interacting partners of PknB and loss of their function resulted in deregulation of central carbon metabolism, with carbon flux diverted to synthesis of the antibiotic actinorhodin.The results indicate that inactivation of FHA 'brake' proteins can potentially amplify the function of STPKs and, in this case, provide a means to overproduce antibiotics.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Science, Swansea University, Singleton Park, Swansea SA28PP, UK.

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Detection of PknB in cellular fractions of E. coli. PknB was detected with HRP conjugated anti penta – His antibody. Lane 1 = whole‐cell extract (WCE) from uninduced E. coli transformed with pLK1. Lane 2 = WCE from induced cells containing pLK1. Lane 3 = insoluble fraction collected after centrifugation of WCE. Lane 4 = soluble fraction. Lane 5 = membrane fraction. Equal amounts of total protein were added to each lane.
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f3: Detection of PknB in cellular fractions of E. coli. PknB was detected with HRP conjugated anti penta – His antibody. Lane 1 = whole‐cell extract (WCE) from uninduced E. coli transformed with pLK1. Lane 2 = WCE from induced cells containing pLK1. Lane 3 = insoluble fraction collected after centrifugation of WCE. Lane 4 = soluble fraction. Lane 5 = membrane fraction. Equal amounts of total protein were added to each lane.

Mentions: PknB is predicted to be a 673‐amino‐acid membrane protein with the ability to autophosphorylate. To investigate these properties, recombinant N‐terminal His‐tagged PknB was expressed in E. coli BL21. After fractionation and Western blotting with an antibody to detect the His‐tagged protein, PknB was predominantly found to be enriched in membrane fractions from E. coli (Fig. 3), migrating with an apparent approximate molecular mass of 130 kDa.


Forkhead-associated proteins genetically linked to the serine/threonine kinase PknB regulate carbon flux towards antibiotic biosynthesis in Streptomyces coelicolor.

Jones G, Del Sol R, Dudley E, Dyson P - Microb Biotechnol (2010)

Detection of PknB in cellular fractions of E. coli. PknB was detected with HRP conjugated anti penta – His antibody. Lane 1 = whole‐cell extract (WCE) from uninduced E. coli transformed with pLK1. Lane 2 = WCE from induced cells containing pLK1. Lane 3 = insoluble fraction collected after centrifugation of WCE. Lane 4 = soluble fraction. Lane 5 = membrane fraction. Equal amounts of total protein were added to each lane.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3818866&req=5

f3: Detection of PknB in cellular fractions of E. coli. PknB was detected with HRP conjugated anti penta – His antibody. Lane 1 = whole‐cell extract (WCE) from uninduced E. coli transformed with pLK1. Lane 2 = WCE from induced cells containing pLK1. Lane 3 = insoluble fraction collected after centrifugation of WCE. Lane 4 = soluble fraction. Lane 5 = membrane fraction. Equal amounts of total protein were added to each lane.
Mentions: PknB is predicted to be a 673‐amino‐acid membrane protein with the ability to autophosphorylate. To investigate these properties, recombinant N‐terminal His‐tagged PknB was expressed in E. coli BL21. After fractionation and Western blotting with an antibody to detect the His‐tagged protein, PknB was predominantly found to be enriched in membrane fractions from E. coli (Fig. 3), migrating with an apparent approximate molecular mass of 130 kDa.

Bottom Line: Here we report functional analysis of pknB and two linked genes, fhaAB, encoding forkhead-associated (FHA) domain proteins that are part of a highly conserved gene locus in actinobacteria.FhaAB are candidate interacting partners of PknB and loss of their function resulted in deregulation of central carbon metabolism, with carbon flux diverted to synthesis of the antibiotic actinorhodin.The results indicate that inactivation of FHA 'brake' proteins can potentially amplify the function of STPKs and, in this case, provide a means to overproduce antibiotics.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Science, Swansea University, Singleton Park, Swansea SA28PP, UK.

Show MeSH
Related in: MedlinePlus