A catalog of associations between rare coding variants and COVID-19 outcomes.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) causes coronavirus disease-19 (COVID-19), a respiratory illness that can result in hospitalization or death. We investigated associations between rare genetic variants and seven COVID-19 outcomes in 543,213 individuals, including 8,248 with COVID-19. After accounting for multiple testing, we did not identify any clear associations with rare variants either exome-wide or when specifically focusing on (i) 14 interferon pathway genes in which rare deleterious variants have been reported in severe COVID-19 patients; (ii) 167 genes located in COVID-19 GWAS risk loci; or (iii) 32 additional genes of immunologic relevance and/or therapeutic potential. Our analyses indicate there are no significant associations with rare protein-coding variants with detectable effect sizes at our current sample sizes. Analyses will be updated as additional data become available, with results publicly browsable at https://rgc-covid19.regeneron.com.

Precise mapping of quantitative trait loci for resistance to southern leaf blight, caused by Cochliobolus heterostrophus race O, and flowering time using advanced intercross maize lines.

The intermated B73 x Mo17 (IBM) population, an advanced intercross recombinant inbred line population derived from a cross between the maize lines B73 (susceptible) and Mo17 (resistant), was evaluated in four environments for resistance to southern leaf blight (SLB) disease caused by Cochliobolus heterostrophus race O. Two environments were artificially inoculated, while two were not inoculated and consequently had substantially lower disease pressure. Four common SLB resistance quantitative trait loci (QTL) were identified in all environments, two in bin 3.04 and one each in bins 1.10 and 8.02/3. There was no significant correlation between disease resistance and days to anthesis. A direct comparison was made between SLB QTL detected in two populations, independently derived from the same parental cross: the IBM advanced intercross population and a conventional recombinant inbred line population. Several QTL for SLB resistance were detected in both populations, with the IBM providing between 5 and, in one case, 50 times greater mapping resolution.

QTL mapping with near-isogenic lines in maize.

A set of 89 near-isogenic lines (NILs) of maize was created using marker-assisted selection. Nineteen genomic regions, identified by restriction fragment length polymorphism loci and chosen to represent portions of all ten maize chromosomes, were introgressed by backcrossing three generations from donor line Tx303 into the B73 genetic background. NILs were genotyped at an additional 128 simple sequence repeat loci to estimate the size of introgressions and the amount of background introgression. Tx303 introgressions ranged in size from 10 to 150 cM, with an average of 60 cM. Across all NILs, 89% of the Tx303 genome is represented in targeted and background introgressions. The average proportion of background introgression was 2.5% (range 0-15%), significantly lower than the expected value of 9.4% for third backcross generation lines developed without marker-assisted selection. The NILs were grown in replicated field evaluations in two years to map QTLs for flowering time traits. A parallel experiment of testcrosses of each NIL to the unrelated inbred, Mo17, was conducted in the same environments to map QTLs in NIL testcross hybrids. QTLs affecting days to anthesis, days to silking, and anthesis-silk interval were detected in both inbreds and hybrids in both environments. The testing environments differed dramatically for drought stress, and different sets of QTLs were detected across environments. Furthermore, QTLs detected in inbreds were typically different from QTLs detected in hybrids, demonstrating the genetic complexity of flowering time. NILs can serve as a valuable genetic mapping resource for maize breeders and geneticists.

Association analysis of candidate genes for maysin and chlorogenic acid accumulation in maize silks.

Two compounds, the C-glycosyl flavone maysin and the phenylpropanoid product chlorogenic acid (CGA), have been implicated in corn earworm (Helicoverpa zea Boddie) resistance in maize (Zea mays L.). Previous quantitative trait locus (QTL) analyses identified the pericarp color (p) locus, which encodes a transcription factor, as the major QTL for maysin and CGA. QTL analysis has also implicated the dihydroflavanol reductase (DFR; E.C. no. 1.1.1.219) locus anthocyaninless1 (a1) and the duplicate chalcone synthase (CHS; E.C. no. 2.3.1.74) loci colorless2 (c2) and white pollen1 (whp1) as genes underlying QTL for maysin and/or CGA synthesis. Epistatic interactions between p and a1 and between p and c2 were also defined. CHS catalyzes the first step in the flavonoid pathway and represents one of the first enzyme steps following the branch off the general phenylpropanoid pathway towards CGA synthesis. In maize, the reduction of dihydroflavanol to leucoanthocyanin by DFR immediately follows the pathway branch leading to C-glycosyl flavone production. The detection of QTLs for maysin and CGA concentration at loci encoding enzyme steps following the pathway branch points implicates alterations in the flow of biochemical intermediates as the biological basis of the QTL effects. To examine if sequence variation among alleles of a1, c2, and whp1 affect maysin and CGA synthesis in maize silks, we performed an association analysis. Because the p locus has often been a major QTL for maysin and CGA and has exhibited epistatic interactions with a1, c2, and whp1, association analysis was conditioned on the p genotype. A highly significant association of two sequence polymorphisms in the promoter of a1 with maysin synthesis was demonstrated. Additional conditioning on the genotype of the significant a1 polymorphism allowed the detection of a significant polymorphism within the whp1 promoter. Our analyses demonstrate that conditioning for epistatic factors greatly increases the power of association testing.

New features and enhancements in the X-PLOR computer program.

This article describes new methods for X-ray crystallographic refinement and nuclear magnetic resonance (NMR) structure determination that are available in the recent release of the X-PLOR software, X-PLOR 98.0. The major new features of the X-PLOR 98.0 software are: (i) the introduction of maximum likelihood methods (Pannu and Read, Acta Crystallogr 1996;A52:659-668) for X-ray crystallographic refinement with structure factor amplitude, intensity and phase probability targets, (ii) the addition of the Andersen thermal coupling method for temperature control during simulated annealing refinements, (iii) a new utility function for converting reflection data in to the X-PLOR format, (iv) validated scripts and performance enhancements for structure determination from NMR distance restraints using torsion angle dynamics, (v) fast code for direct nuclear Oberhauser effect (NOE) refinement using matrix doubling and gaussian quadratures, (vi) methodologies for using ambiguous restraint information to perform automated iterative peak assignment and structure determination (Nilges et al., J Mol Biol 1997;269: 408-422). Additional developments in methodology for refining crystal structures from poor initial models include the implementation of a fast adaptive bulk solvent scattering correction and an energy minimization routine that makes use of second derivative information. Trial crystallographic refinements with an energy minimization protocol that includes these enhancements indicate significantly improved convergence. The quality of the resulting models appears comparable to models obtained from refinement protocols that incorporate torsion angle dynamics. Test applications of the new energy minimizer to NMR structure refinement with using NOE calculations also show improved convergence, leading to more optimized final models.

Determination of (3)J(H (infi) (supN) ,C (infi) (sup') ) coupling constants in proteins with the C'-FIDS method.

We introduce the C'-FIDS-(1)H,(15)N-HSQC experiment, a new method for the determination of (3)J(H (infi) (supN) ,C (infi) (sup') ) coupling constants in proteins, yielding information about the torsional angle ϕ. It relies on the (1)H,(15)N-HSQC or HNCO experiment, two of the the most sensitive heteronuclear correlation experiments for isotopically labeled proteins. A set of three (1)H,(15)N-HSQC or HNCO spectra are recorded: a reference experiment in which the carbonyl spins are decoupled during t(1) and t(2), a second experiment in which they are decoupled exclusively during t(1) and a third one in which they are coupled in t(1) as well as t(2). The last experiment yields an E.COSY-type pattern from which the (2)J(H (infi) (supN) ,C (infi-1) (sup') ) and (1)J(N(i),C (infi-1) (sup') ) coupling constants can be extracted. By comparison of the coupled multiplet (obtained from the second experiment) with the decoupled multiplet (obtained from the first experiment) convoluted with the (2)J(H (infi) (supN) ,C (infi-1) (sup') ) coupling, the (3)J(H (infi) (supN) ,C (infi) (sup') ) coupling can be found in a one-parameter fitting procedure. The method is demonstrated for the protein rhodniin, containing 103 amino acids. Systematic errors due to differential relaxation are small for (n)J(H(N),C') couplings in biomacromolecules of the size currently under NMR spectroscopic investigation.