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Summary This work revisits a publication by Beanet al.(2018) that reports seven amino acid substitutions are essential for the evolution ofl‐DOPA 4,5‐dioxygenase (DODA) activity in Caryophyllales. In this study, we explore several concerns which led us to replicate the analyses of Beanet al.(2018).Our comparative analyses, with structural modelling, implicate numerous residues additional to those identified by Beanet al.(2018), with many of these additional residues occurring around the active site of BvDODAα1. We therefore replicated the analyses of Beanet al.(2018) to re‐observe the effect of their original seven residue substitutions in a BvDODAα2 background, that is the BvDODAα2‐mut3 variant.Multiplein vivoassays, in bothSaccharomyces cerevisiaeandNicotiana benthamiana, did not result in visible DODA activity in BvDODAα2‐mut3, with betalain production always 10‐fold below BvDODAα1.In vitroassays also revealed substantial differences in both catalytic activity and pH optima between BvDODAα1, BvDODAα2 and BvDODAα2‐mut3 proteins, explaining their differing performancein vivo.In summary, we were unable to replicate thein vivoanalyses of Beanet al.(2018), and our quantitativein vivoandin vitroanalyses suggest a minimal effect of these seven residues in altering catalytic activity of BvDODAα2. We conclude that the evolutionary pathway to high DODA activity is substantially more complex than implied by Beanet al.(2018).
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One-Step Replica Symmetry Breaking of Random Regular NAE-SAT II
Abstract Continuing our earlier work in Nam et al. (One-step replica symmetry breaking of random regular NAE-SAT I,arXiv:2011.14270, 2020), we study the random regulark-nae-satmodel in the condensation regime. In Nam et al. (2020), the (1rsb) properties of the model were established with positive probability. In this paper, we improve the result to probability arbitrarily close to one. To do so, we introduce a new framework which is the synthesis of two approaches: the small subgraph conditioning and a variance decomposition technique using Doob martingales and discrete Fourier analysis. The main challenge is a delicate integration of the two methods to overcome the difficulty arising from applying the moment method to an unbounded state space.
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- Award ID(s):
- 1954337
- PAR ID:
- 10492211
- Publisher / Repository:
- Springer Science + Business Media
- Date Published:
- Journal Name:
- Communications in Mathematical Physics
- Volume:
- 405
- Issue:
- 3
- ISSN:
- 0010-3616
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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