Outer membrane protein genes and their small non-coding RNA regulator genes in Photorhabdus luminescens

INTRODUCTION: Three major outer membrane protein genes of Escherichia coli, ompF, ompC, and ompA respond to stress factors. Transcripts from these genes are regulated by the small non-coding RNAs micF, micC, and micA, respectively. Here we examine Photorhabdus luminescens, an organism that has a different habitat from E. coli for outer membrane protein genes and their regulatory RNA genes. RESULTS: By bioinformatics analysis of conserved genetic loci, mRNA 5'UTR sequences, RNA secondary structure motifs, upstream promoter regions and protein sequence homologies, an ompF -like porin gene in P. luminescens as well as a duplication of this gene have been predicted. Gene loci for micF RNA, as well as OmpC protein and its associated regulatory micC RNA, were not found. Significantly, a sequence bearing the appropriate signatures of the E. coli micA RNA was located. The ompA homolog was previously annotated in P. luminescens. CONCLUSION: Presence of an ompF-like porin in P. luminescens is in keeping with the necessity to allow for passage of small molecules into the cell. The apparent lack of ompC, micC and micF suggests that these genes are not essential to P. luminescens and ompC and micF in particular may have been lost when the organism entered its defined life cycle and partially protected habitat. Control of porin gene expression by RNA may be more prevalent in free- living cells where survival is dependent on the ability to make rapid adjustments in response to environmental stress. Regulation of ompA by micA may have been retained due to a necessity for ompA control during one or both stages of the P. luminescens life cycle. REVIEWERS: This article was reviewed by Tal Dagan (nominated by Dan Graur), Mikhail Gelfand and Anna Gerasimova (nominated by Mikhail Gelfand) and J Peter Gogarten. OPEN PEER REVIEW: Reviewed by Tal Dagan (nominated by Dan Graur), Mikhail Gelfand and Anna Gerasimova (nominated by Mikhail Gelfand) and J Peter Gogarten. For the full reviews, please go to the Reviewers' comments section

Synthesis Cost-Optimal Targeted Mutant Protein Libraries

Protein variant libraries produced by site-directed mutagenesis are a useful tool utilized by protein engineers to explore variants with potentially improved properties, such as activity and stability. These libraries are commonly built by selecting residue positions and alternative beneficial mutations for each position. All possible combinations are then constructed and screened, by incorporating degenerate codons at mutation sites. These degenerate codons often encode additional unwanted amino acids or even STOP codons. Our study aims to take advantage of annealing based recombination of oligonucleotides during synthesis and utilize multiple degenerate codons per mutation site to produce targeted protein libraries devoid of unwanted variants. Toward this goal we created an algorithm to calculate the minimum number of degenerate codons necessary to specify any given amino acid set, and a dynamic programming method that uses this algorithm to optimally partition a DNA target sequence with degeneracies into overlapping oligonucleotides, such that the total cost of synthesis of the target mutant protein library is minimized. Computational experiments show that, for a modest increase in DNA synthesis costs, beneficial variant yields in produced mutant libraries are increased by orders of magnitude, an effect particularly pronounced in large combinatorial libraries

Improved Algorithms for Approximate String Matching (Extended Abstract)

The problem of approximate string matching is important in many different areas such as computational biology, text processing and pattern recognition. A great effort has been made to design efficient algorithms addressing several variants of the problem, including comparison of two strings, approximate pattern identification in a string or calculation of the longest common subsequence that two strings share. We designed an output sensitive algorithm solving the edit distance problem between two strings of lengths n and m respectively in time O((s-|n-m|)min(m,n,s)+m+n) and linear space, where s is the edit distance between the two strings. This worst-case time bound sets the quadratic factor of the algorithm independent of the longest string length and improves existing theoretical bounds for this problem. The implementation of our algorithm excels also in practice, especially in cases where the two strings compared differ significantly in length. Source code of our algorithm is available at http://www.cs.miami.edu/\~dimitris/edit_distanceComment: 10 page

High-resolution global grids of revised Priestley–Taylor and Hargreaves–Samani coefficients for assessing ASCE-standardized reference crop evapotranspiration and solar radiation

Abstract. The objective of the study is to provide global grids (0.5°) of revised annual coefficients for the Priestley–Taylor (P-T) and Hargreaves–Samani (H-S) evapotranspiration methods after calibration based on the ASCE (American Society of Civil Engineers)-standardized Penman–Monteith method (the ASCE method includes two reference crops: short-clipped grass and tall alfalfa). The analysis also includes the development of a global grid of revised annual coefficients for solar radiation (Rs) estimations using the respective Rs formula of H-S. The analysis was based on global gridded climatic data of the period 1950–2000. The method for deriving annual coefficients of the P-T and H-S methods was based on partial weighted averages (PWAs) of their mean monthly values. This method estimates the annual values considering the amplitude of the parameter under investigation (ETo and Rs) giving more weight to the monthly coefficients of the months with higher ETo values (or Rs values for the case of the H-S radiation formula). The method also eliminates the effect of unreasonably high or low monthly coefficients that may occur during periods where ETo and Rs fall below a specific threshold. The new coefficients were validated based on data from 140 stations located in various climatic zones of the USA and Australia with expanded observations up to 2016. The validation procedure for ETo estimations of the short reference crop showed that the P-T and H-S methods with the new revised coefficients outperformed the standard methods reducing the estimated root mean square error (RMSE) in ETo values by 40 and 25 %, respectively. The estimations of Rs using the H-S formula with revised coefficients reduced the RMSE by 28 % in comparison to the standard H-S formula. Finally, a raster database was built consisting of (a) global maps for the mean monthly ETo values estimated by ASCE-standardized method for both reference crops, (b) global maps for the revised annual coefficients of the P-T and H-S evapotranspiration methods for both reference crops and a global map for the revised annual coefficient of the H-S radiation formula and (c) global maps that indicate the optimum locations for using the standard P-T and H-S methods and their possible annual errors based on reference values. The database can support estimations of ETo and solar radiation for locations where climatic data are limited and it can support studies which require such estimations on larger scales (e.g. country, continent, world). The datasets produced in this study are archived in the PANGAEA database (https://doi.org/10.1594/PANGAEA.868808) and in the ESRN database (http://www.esrn-database.org or http://esrn-database.weebly.com)