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- Gaca, Anthony O., et al.
(författare)
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From (p)ppGpp to (pp)pGpp : characterization of Regulatory Effects of pGpp Synthesized by the Small Alarmone Synthetase of Enterococcus faecalis
- 2015
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Ingår i: Journal of Bacteriology. - : American Society for Microbiology. - 0021-9193 .- 1098-5530. ; 197:18, s. 2908-2919
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Tidskriftsartikel (refereegranskat)abstract
- The bacterial stringent response (SR) is a conserved stress tolerance mechanism that orchestrates physiological alterations to enhance cell survival. This response is mediated by the intracellular accumulation of the alarmones pppGpp and ppGpp, collectively called (p) ppGpp. In Enterococcus faecalis, (p) ppGpp metabolism is carried out by the bifunctional synthetase/hydrolase E. faecalis Rel (Rel(Ef)) and the small alarmone synthetase (SAS) RelQ(Ef). Although Rel is the main enzyme responsible for SR activation in Firmicutes, there is emerging evidence that SASs can make important contributions to bacterial homeostasis. Here, we showed that RelQ(Ef) synthesizes ppGpp more efficiently than pppGpp without the need for ribosomes, tRNA, or mRNA. In addition to (p) ppGpp synthesis from GDP and GTP, RelQ(Ef) also efficiently utilized GMP to form GMP 3'-diphosphate (pGpp). Based on this observation, we sought to determine if pGpp exerts regulatory effects on cellular processes affected by (p) ppGpp. We found that pGpp, like (p) ppGpp, strongly inhibits the activity of E. faecalis enzymes involved in GTP biosynthesis and, to a lesser extent, transcription of rrnB by Escherichia coli RNA polymerase. Activation of E. coli RelA synthetase activity was observed in the presence of both pGpp and ppGpp, while RelQ(Ef) was activated only by ppGpp. Furthermore, enzymatic activity of RelQ(Ef) is insensitive to relacin, a (p) ppGpp analog developed as an inhibitor of "long" RelA/SpoT homolog (RSH) enzymes. We conclude that pGpp can likely function as a bacterial alarmone with target-specific regulatory effects that are similar to what has been observed for (p) ppGpp. IMPORTANCE Accumulation of the nucleotide second messengers (p) ppGpp in bacteria is an important signal regulating genetic and physiological networks contributing to stress tolerance, antibiotic persistence, and virulence. Understanding the function and regulation of the enzymes involved in (p) ppGpp turnover is therefore critical for designing strategies to eliminate the protective effects of this molecule. While characterizing the (p) ppGpp synthetase RelQ of Enterococcus faecalis (RelQ(Ef)), we found that, in addition to (p) ppGpp, RelQ(Ef) is an efficient producer of pGpp (GMP 3'-diphosphate). In vitro analysis revealed that pGpp exerts complex, target-specific effects on processes known to be modulated by (p) ppGpp. These findings provide a new regulatory feature of RelQ(Ef) and suggest that pGpp may represent a new member of the (pp) pGpp family of alarmones.
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| 2. |
- Wang, Kun
(författare)
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Migration Towards Next Generation Optical Access and Transport Networks
- 2017
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- By 2020 there will be 50 billion connected devices over the Internet. With the fast-increasing data traffic demand in both fixed and mobile networks, network operators need to migrate networks towards next generation solutions. The network migration requires the enormous investment in equipment and infrastructure, while the revenues are not expected to grow significantly. Therefore, one of the main challenges for network operators is to find out a proper cost-effective optical network solution that can match future high capacity demand and flexibly support multiple network services on a common network infrastructure.The first part of the thesis addresses the Active Optical Network (AON) and its migration strategies towards Next Generation Optical Access (NGOA) solutions. Several migration strategies are proposed from the perspective of network topology, data plane and control plane. A general methodology for Techno-Economic analysis has been developed and applied to the Total Cost of Ownership (TCO) calculation of different NGOA solutions. The thesis provides a complete cost evaluation of AON migration paths, which can be used by network operators to assess the economic feasibility of network migration.A converged Optical Transport Network (OTN) that can serve both fixed and mobile network services is beneficial from the cost-saving perspective. However, the different types of services, require different network performance. The second part of the thesis focuses on the investigation of the converged OTN that can be flexibly and timely adjusted to satisfy varying service conditions. A programmable OTN featured with Wavelength Division Multiplexing (WDM) in the data plane and Software Defined Networking (SDN) in control plane has been proposed. To demonstrate the benefits of the converged OTN, the thesis also provides a multi-domain orchestration architecture for the multiple network services. The resource orchestration, across three network domains: OTN, mobile network and cloud, enables agile service creation and optimized resource allocation among the multiple domains.
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