Swap Experiment Research Articles

Mutagenesis studies of TRPV1 subunit interfaces informed by genomic variant analysis

Protein structures and mutagenesis studies have been instrumental in elucidating molecular mechanisms of ion channel function, but making informed choices about which residues to target for mutagenesis can be challenging. Therefore, we investigated the potential for using human population genomic data to further refine our selection of mutagenesis sites in TRPV1. Single nucleotide polymorphism data of TRPV1 from gnomAD 2.1.1 revealed a lower number of missense variants within buried residues of the ankyrin repeat domain and an increased number of variants between secondary structure elements of the transmembrane segments. We hypothesized that residues critical to interactions at interfaces between subunits or domains in the channel would exhibit a similar reduction in variants. We identified in the structure of ground squirrel TRPV1 (PDB: 7LQY) a possible electrostatic network between K155 and K160 in the N-terminal ankyrin repeat domain and E761 and D762 in the C-terminus (K-KED). Consistent with our hypothesis for residues at key interface sites, none of the four residues have any variants reported in gnomAD 2.1.1. Ca2+ imaging of TRPV1 K-KED mutants confirmed significant roles for these residues, but we found that the electrostatic interaction is not essential since channel function is still observed in total charge reversals on the C-terminal side of the interface (E761K/D762K). Interestingly, Ca2+ imaging responses for a charge swap experiment with K155D/D762K showed partially restored wild-type responses. Using electrophysiology, we found that charge reversals on either K155 or D762 increased the baseline currents of TRPV1, and the charge swapped double mutant, K155D/D762K, partially restored baseline currents to wild-type levels. We interpret these results to mean that contacts across residues in the K-KED interface shift the equilibria of conformations to closed pore states. Our study demonstrates the utility and applicability of a combined missense variant and structure targeted investigation of residues at TRPV1 subunit interfaces.

MtEFD and MtEFD2: Two transcription factors with distinct neofunctionalization in symbiotic nodule development.

Rhizobium-legume nitrogen-fixing symbiosis involves the formation of a specific organ, the root nodule, which provides bacteria with the proper cellular environment for atmospheric nitrogen fixation. Coordinated differentiation of plant and bacterial cells is an essential step of nodule development, for which few transcriptional regulators have been characterized. Medicago truncatula ETHYLENE RESPONSE FACTOR REQUIRED FOR NODULE DIFFERENTIATION (MtEFD) encodes an APETALA2/ETHYLENE RESPONSIVE FACTOR (ERF) transcription factor, the mutation of which leads to both hypernodulation and severe defects in nodule development. MtEFD positively controls a negative regulator of cytokinin signaling, the RESPONSE REGULATOR 4 (MtRR4) gene. Here we showed that that the Mtefd-1 mutation affects both plant and bacterial endoreduplication in nodules, as well as the expression of hundreds of genes in young and mature nodules, upstream of known regulators of symbiotic differentiation. MtRR4 expressed with the MtEFD promoter complemented Mtefd-1 hypernodulation but not the nodule differentiation phenotype. Unexpectedly, a nonlegume homolog of MtEFD, AtERF003 in Arabidopsis (Arabidopsis thaliana), could efficiently complement both phenotypes of Mtefd-1, in contrast to the MtEFD paralog MtEFD2 expressed in the root and nodule meristematic zone. A domain swap experiment showed that MtEFD2 differs from MtEFD by its C-terminal fraction outside the DNA binding domain. Furthermore, clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9 (CRISPR-Cas9)mutagenesis of MtEFD2 led to a reduction in the number of nodules formed in Mtefd-1, with downregulation of a set of genes, including notably NUCLEAR FACTOR-YA1 (MtNF-YA1) and MtNF-YB16, which are essential for nodule meristem establishment. We, therefore, conclude that nitrogen-fixing symbiosis recruited two proteins originally expressed in roots, MtEFD and MtEFD2, with distinct functions and neofunctionalization processes for each of them.

Influence of Plant Fraction, Soil, and Plant Species on Microbiota: a Multikingdom Comparison.

Plant roots influence the soil microbiota via physical interaction, secretion, and plant immunity. However, it is unclear whether the root fraction or soil is more important in determining the structure of the prokaryotic or eukaryotic community and whether this varies between plant species. Furthermore, the leaf (phyllosphere) and root microbiotas have a large overlap; however, it is unclear whether this results from colonization of the phyllosphere by the root microbiota. Soil, rhizosphere, rhizoplane, and root endosphere prokaryote-, eukaryote-, and fungus-specific microbiotas of four plant species were analyzed with high-throughput sequencing. The strengths of factors controlling microbiota structure were determined using permutational multivariate analysis of variance (PERMANOVA) statistics. The origin of the phyllosphere microbiota was investigated using a soil swap experiment. Global microbial kingdom analysis conducted simultaneously on multiple plants shows that cereals, legumes, and Brassicaceae establish similar prokaryotic and similar eukaryotic communities inside and on the root surface. While the bacterial microbiota is recruited from the surrounding soil, its profile is influenced by the root itself more so than by soil or plant species. However, in contrast, the fungal microbiota is most strongly influenced by soil. This was observed in two different soils and for all plant species examined. Microbiota structure is established within 2 weeks of plant growth in soil and remains stable thereafter. A reciprocal soil swap experiment shows that the phyllosphere is colonized from the soil in which the plant is grown.IMPORTANCE Global microbial kingdom analysis conducted simultaneously on multiple plants shows that cereals, legumes, and Brassicaceae establish similar prokaryotic and similar eukaryotic communities inside and on the root surface. While the bacterial microbiota is recruited from the surrounding soil, its profile is influenced by the root fraction more so than by soil or plant species. However, in contrast, the fungal microbiota is most strongly influenced by soil. This was observed in two different soils and for all plant species examined, indicating conserved adaptation of microbial communities to plants. Microbiota structure is established within 2 weeks of plant growth in soil and remains stable thereafter. We observed a remarkable similarity in the structure of a plant's phyllosphere and root microbiotas and show by reciprocal soil swap experiments that both fractions are colonized from the soil in which the plant is grown. Thus, the phyllosphere is continuously colonized by the soil microbiota.

The AtMYB12 activation domain maps to a short C-terminal region of the transcription factor.

The Arabidopsis thaliana R2R3-MYB transcription factor MYB12 is a light-inducible, flavonol-specific activator of flavonoid biosynthesis. The transactivation activity of the AtMYB12 protein was analyzed using a C-terminal deletion series in a transient A. thaliana protoplast assay with the goal of mapping the activation domain (AD). Although the deletion of the last 46 C-terminal amino acids did not affect the activation capacity, the deletion of the last 98 amino acids almost totally abolished transactivation of two different target promoters. A domain swap experiment using the yeast GAL4 DNA-binding domain revealed that the region from positions 282 to 328 of AtMYB12 was sufficient for transactivation. In contrast to the R2R3-MYB ADs known thus far, that of AtMYB12 is not located at the rearmost C-terminal end of the protein. The AtMYB12 AD is conserved in other experimentally proven R2R3-MYB flavonol regulators from different species.

Relative importance of soil properties and microbial community for soil functionality: insights from a microbial swap experiment

Summary The most accepted theories in soil ecology suggest that broad (e.g. respiration) and specialized (e.g. denitrification) functions are affected differently by resource availability and microbial communities in terrestrial ecosystems. However, there is a lack of experimental approaches quantifying and separating the role of microbial communities from the effect of soil abiotic properties on different aspects of soil ecosystem functionality. Here, we conducted a full‐factorial design microcosm experiment and used random forest and structural equation modelling (SEM) analyses to evaluate the role and the relative importance of soil properties (sterile soils A, B and C differing in abiotic attributes) and microbial communities (microbial inoculums from soils A, B and C) in driving soil respiration (i.e. broad functioning), denitrification (i.e. specialized functioning) and four enzyme activities and carbon (C) and nitrogen (N) availability in soil. Soils with the higher total C (soils B and C) promoted the highest soil C and N availability, enzyme activities and broad functioning (i.e. soil respiration); however, we did not find any effect of total C on specialized functions (i.e. denitrification rates). Random forest analyses showed that both soil properties (i.e. total C and pH) and microbial abundance determined broad functioning (i.e. soil respiration), as well as the production of enzyme activities, and C and N availability in soil. However, we found that microbial communities were more important than soil properties for modulating specialized functioning (i.e. denitrification rates) in soil environments. Finally, our SEM also indicated that broad functioning, which is widely distributed across living organisms, is limited by both resource availability and microbial abundance. Furthermore, specialized functioning, which is conducted by particular groups of organisms, may be highly sensitive to changes in the microbial community. Overall, our findings provide direct experimental evidence for the relative importance of soil properties and microbial communities on broad and specialized functioning. Such evidence helps advance our understanding of different drivers of soil ecosystem functioning which will be crucial to developing an ecologically relevant theory about below‐ground ecosystem functioning. A lay summary is available for this article.

Two Predicted Transmembrane Domains Exclude Very Long Chain Fatty acyl-CoAs from the Active Site of Mouse Wax Synthase.

Wax esters are used as coatings or storage lipids in all kingdoms of life. They are synthesized from a fatty alcohol and an acyl-CoA by wax synthases. In order to get insights into the structure-function relationships of a wax synthase from Mus musculus, a domain swap experiment between the mouse acyl-CoA:wax alcohol acyltransferase (AWAT2) and the homologous mouse acyl-CoA:diacylglycerol O-acyltransferase 2 (DGAT2) was performed. This showed that the substrate specificity of AWAT2 is partially determined by two predicted transmembrane domains near the amino terminus of AWAT2. Upon exchange of the two domains for the respective part of DGAT2, the resulting chimeric enzyme was capable of incorporating up to 20% of very long acyl chains in the wax esters upon expression in S. cerevisiae strain H1246. The amount of very long acyl chains in wax esters synthesized by wild type AWAT2 was negligible. The effect was narrowed down to a single amino acid position within one of the predicted membrane domains, the AWAT2 N36R variant. Taken together, we provide first evidence that two predicted transmembrane domains in AWAT2 are involved in determining its acyl chain length specificity.

Identification of a Novel Protein Binding Motif within the T-synthase for the Molecular Chaperone Cosmc

Prior studies suggested that the core 1 β3-galactosyltransferase (T-synthase) is a specific client of the endoplasmic reticulum chaperone Cosmc, whose function is required for T-synthase folding, activity, and consequent synthesis of normal O-glycans in all vertebrate cells. To explore whether the T-synthase encodes a specific recognition motif for Cosmc, we used deletion mutagenesis to identify a cryptic linear and relatively hydrophobic peptide in the N-terminal stem region of the T-synthase that is essential for binding to Cosmc (Cosmc binding region within T-synthase, or CBRT). Using this sequence information, we synthesized a peptide containing CBRT and found that it directly interacts with Cosmc and also inhibits Cosmc-assisted in vitro refolding of denatured T-synthase. Moreover, engineered T-synthase carrying mutations within CBRT exhibited diminished binding to Cosmc that resulted in the formation of inactive T-synthase. To confirm the general recognition of CBRT by Cosmc, we performed a domain swap experiment in which we inserted the stem region of the T-synthase into the human β4GalT1 and found that the CBRT element can confer Cosmc binding onto the β4GalT1 chimera. Thus, CBRT is a unique recognition motif for Cosmc to promote its regulation and formation of active T-synthase and represents the first sequence-specific chaperone recognition system in the ER/Golgi required for normal protein O-glycosylation.

Constitutive and Nitrogen Catabolite Repression-sensitive Production of Gat1 Isoforms

Nitrogen catabolite repression (NCR)-sensitive transcription is activated by Gln3 and Gat1. In nitrogen excess, Gln3 and Gat1 are cytoplasmic, and transcription is minimal. In poor nitrogen, Gln3 and Gat1 become nuclear and activate transcription. A long standing paradox has surrounded Gat1 production. Gat1 was first reported as an NCR-regulated activity mediating NCR-sensitive transcription in gln3 deletion strains. Upon cloning, GAT1 transcription was, as predicted, NCR-sensitive and Gln3- and Gat1-activated. In contrast, Western blots of Gat1-Myc(13) exhibited two constitutively produced species. Investigating this paradox, we demonstrate that wild type Gat1 isoforms (IsoA and IsoB) are initiated at Gat1 methionines 40, 95, and/or 102, but not at methionine 1. Their low level production is the same in rich and poor nitrogen conditions. When the Myc(13) tag is placed after Gat1 Ser-233, four N-terminal Gat1 isoforms (IsoC-F) are also initiated at methionines 40, 95, and/or 102. However, their production is highly NCR-sensitive, being greater in proline than glutamine medium. Surprisingly, all Gat1 isoforms produced in sufficient quantities to be confidently analyzed (IsoA, IsoC, and IsoD) require Gln3 and UASGATA promoter elements, both requirements typical of NCR-sensitive transcription. These data demonstrate that regulated Gat1 production is more complex than previously recognized, with wild type versus truncated Gat1 proteins failing to be regulated in parallel. This is the first reported instance of Gln3 UASGATA-dependent protein production failing to derepress in nitrogen poor conditions. A Gat1-lacZ ORF swap experiment indicated sequence(s) responsible for the nonparallel production are downstream of Gat1 leucine 61.

Acceptance by Honey Bee Guards of Non-Nestmates is not Increased by Treatment with Nestmate Odours

Honey bee, Apis mellifera, entrance guards use chemical cues to discriminate nestmates from non-nestmates. Previous research has shown that when wax combs are reciprocally swapped between two colonies, guards become more accepting of workers from the swap partner. However, when combs were transferred only one way, guards in the comb-receiver colony became more accepting of bees from the comb-donor colony, but not vice versa. Hence, the increased acceptance of non-nestmates caused by reciprocal comb swapping was not because of introduced bees acquiring odours from the transferred combs, which was surprising because comb wax was known to affect the odour of bees. In the current experiment, we caused workers to acquire either nestmate or non-nestmate odours by holding them for 15 min in a tube, which had previously held nestmates or non-nestmates and then measured their acceptance by entrance guards of nestmate or non-nestmate hives. When transferred workers had acquired odours of non-nestmates, acceptance by their own colony’s guards significantly decreased to 66% from 91%. Conversely, the acceptance of non-nestmates that had acquired odours of the guards’ own nestmates was unchanged, 25% vs. 25%. These results show that when equivalent changes in the odour of introduced bees are made, guards are more sensitive to changes that cause nestmates to acquire non-nestmate odours than vice versa. These results are also a likely explanation for the earlier and surprising results from the unidirectional comb swap experiment (Couvillon et al. 2007). We make a hypothesis for the underlying mechanism in terms of a multidimensional recognition cue space.

Fertilization in C. elegans requires an intact C-terminal RING finger in sperm protein SPE-42

BackgroundThe C. elegans sperm protein SPE-42, a membrane protein of unknown structure and molecular function, is required for fertilization. Sperm from worms with spe-42 mutations appear normal but are unable to fertilize eggs. Sequence analysis revealed the presence of 8 conserved cysteine residues in the C-terminal cytoplasmic domain of this protein suggesting these residues form a zinc-coordinating RING finger structure.ResultsWe made an in silico structural model of the SPE-42 RING finger domain based on primary sequence analysis and previously reported RING structures. To test the model, we created spe-42 transgenes coding for mutations in each of the 8 cysteine residues predicted to coordinate Zn++ ions in the RING finger motif. Transgenes were crossed into a spe-42 null background and protein function was measured by counting progeny. We found that all 8 cysteines are required for protein function. We also showed that sequence differences between the C-terminal 29 and 30 amino acids in C. elegans and C. briggsae SPE-42 following the RING finger domain are not responsible for the failure of the C. briggsae SPE-42 homolog to rescue C. elegans spe-42 mutants.ConclusionsThe results suggest that a bona fide RING domain is present at the C-terminus of the SPE-42 protein and that this motif is required for sperm-egg interactions during C. elegans fertilization. Our structural model of the RING domain provides a starting point for further structure-function analysis of this critical region of the protein. The C-terminal domain swap experiment suggests that the incompatibility between the C. elegans and C. briggsae SPE-42 proteins is caused by small amino acid differences outside the C-terminal domain.

The Mdm2 RING domain C-terminus is required for supramolecular assembly and ubiquitin ligase activity

Mdm2, a key negative regulator of the p53 tumor suppressor, is a RING-type E3 ubiquitin ligase. The Mdm2 RING domain can be biochemically fractionated into two discrete species, one of which exists as higher order oligomers that are visible by electron microscopy, whereas the other is a monomer. Both fractions are ATP binding and E3 ligase activity competent, although the oligomeric fraction exhibits lower dependence on the E2 component of ubiquitin polymerization reactions. The extreme C-terminal five amino acids of Mdm2 are essential for E3 ligase activity in vivo and in vitro, as well as for oligomeric assembly of the protein. A single residue (phenylalanine 490) in that sequence is critical for both properties. Interestingly, the C-terminus of the Mdm2 homologue, MdmX (itself inert as an E3 ligase), can fully substitute for the equivalent segment of Mdm2 and restore its E3 activity. We further show that the Mdm2 C-terminus is involved in intramolecular interactions and can set up a platform for direct protein-protein interactions with the E2.

Regulated intramembrane proteolysis of FtsL protein and the control of cell division in Bacillus subtilis

The small bitopic division protein FtsL is an essential part of the division machinery (divisome) in most eubacteria. In Bacillus subtilis FtsL is a highly unstable protein and the turnover has been implicated in regulation of division in response to DNA damage. N-terminal deletions and a domain swap experiment identified the short cytoplasmic domain of FtsL as being required for instability. We then identified a zinc metalloprotease, YluC, required for turnover, and likely sequence motifs involved in substrate recognition. YluC belongs to the site-2-protease (S2P) family of proteases involved in regulated intramembrane proteolysis (RIP), which plays a role in diverse regulatory phenomena from bacteria to man. The yluC mutant, and strains with N-terminal truncations of ftsL have a short cell phenotype, indicating that that FtsL is normally rate-limiting for division. Coexpression experiments of FtsL and YluC in Escherichia coli corroborated a model in which FtsL is directly cleaved by the membrane metalloprotease. The results shed new light on the regulation of cell division in B. subtilis and identify a novel class of targets for RIP.

No Evidence For Large-scale Germline Genomic Aberrations in Hereditary Bladder Cancer Patients with High-Resolution Array-Based Comparative Genomic Hybridization

Linkage studies in high-risk families have led to the identification of several important susceptibility genes for hereditary cancer. Unfortunately, such studies offer limited possibilities in the search for high-penetrance bladder cancer genes, as extended bladder cancer families are very rare.

The membrane anchors of the heme chaperone CcmE and the periplasmic thioredoxin CcmG are functionally important

The cytochrome c maturation system of Escherichia coli contains two monotopic membrane proteins with periplasmic, functional domains, the heme chaperone CcmE and the thioredoxin CcmG. We show in a domain swap experiment that the membrane anchors of these proteins can be exchanged without drastic loss of function in cytochrome c maturation. By contrast, the soluble periplasmic forms produced with a cleavable OmpA signal sequence have low biological activity. Both the chimerical CcmE (CcmG′–′E) and the soluble periplasmic CcmE produce low levels of holo-CcmE and thus are impaired in their heme receiving capacity. Also, both forms of CcmE can be co-precipitated with CcmC, thus restricting the site of interaction of CcmE with CcmC to the C-terminal periplasmic domain. However, the low level of holo-CcmE formed in the chimera is transferred efficiently to cytochrome c, indicating that heme delivery from CcmE does not involve the membrane anchor.

Structural Analysis of Arabidopsis thaliana Nucleoside Diphosphate Kinase-2 for Phytochrome-mediated Light Signaling

In plants, nucleoside diphosphate kinases (NDPKs) play a key role in the signaling of both stress and light. However, little is known about the structural elements involved in their function. Of the three NDPKs (NDPK1-NDPK3) expressed in Arabidopsis thaliana, NDPK2 is involved in phytochrome-mediated signal transduction. In this study, we found that the binding of dNDP or NTP to NDPK2 strengthens the interaction significantly between activated phytochrome and NDPK2. To better understand the structural basis of the phytochrome-NDPK2 interaction, we determined the X-ray structures of NDPK1, NDPK2, and dGTP-bound NDPK2 from A.thaliana at 1.8A, 2.6A, and 2.4A, respectively. The structures showed that nucleotide binding caused a slight conformational change in NDPK2 that was confined to helices alphaA and alpha2. This suggests that the presence of nucleotide in the active site and/or the evoked conformational change contributes to the recognition of NDPK2 by activated phytochrome. In vitro binding assays showed that only NDPK2 interacted specifically with the phytochrome and the C-terminal regulatory domain of phytochrome is involved in the interaction. A domain swap experiment between NDPK1 and NDPK2 showed that the variable C-terminal region of NDPK2 is important for the activation by phytochrome. The structure of Arabidopsis NDPK1 and NDPK2 showed that the isoforms share common electrostatic surfaces at the nucleotide-binding site, but the variable C-terminal regions have distinct electrostatic charge distributions. These findings suggest that the binding of nucleotide to NDPK2 plays a regulatory role in phytochrome signaling and that the C-terminal extension of NDPK2 provides a potential binding surface for the specific interaction with phytochromes.

TRNA leucine identity and recognition sets

Transfer RNAs (tRNAs) are grouped into two classes based on the structure of their variable loop. In Escherichia coli, tRNAs from three isoaccepting groups are classified as type II. Leucine tRNAs comprise one such group. We used both in vivo and in vitro approaches to determine the nucleotides that are required for tRNALeu function. In addition, to investigate the role of the tRNA fold, we compared the in vivo and in vitro characteristics of type I tRNALeu variants with their type II counterparts.A minimum of six conserved tRNALeu nucleotides were required to change the amino acid identity and recognition of a type II tRNASer amber suppressor from a serine to a leucine residue. Five of these nucleotides affect tRNA tertiary structure; the G15-C48 tertiary “Levitt base-pair” in tRNASer was changed to A15-U48; the number of nucleotides in the α and β regions of the D-loop was changed to achieve the positioning of G18 and G19 that is found in all tRNALeu; a base was inserted at position 47n between the base-paired extra stem and the T-stem; in addition the G73 “discriminator” base of tRNASer was changed to A73. This minimally altered tRNASer exclusively inserted leucine residues and was an excellent in vitro substrate for LeuRS. In a parallel experiment, nucleotide substitutions were made in a glutamine-inserting type I tRNA (RNASerΔ; an amber suppressor in which the tRNASer type II extra-stem-loop is replaced by a consensus type I loop). This “type I” swap experiment was successful both in vivo and in vitro but required more nucleotide substitutions than did the type II swap.The type I and II swaps revealed differences in the contributions of the tRNALeu acceptor stem base-pairs to tRNALeu function: in the type I, but not the type II fold, leucine specificity was contingent on the presence of the tRNALeu acceptor stem sequence. The type I and II tRNAs used in this study differed only in the sequence and structure of the variable loop. By altering this loop, and thereby possibly introducing subtle changes into the overall tRNA fold, it became possible to detect otherwise cryptic contributions of the acceptor stem sequence to recognition by LeuRS. Possible reasons for this effect are discussed.

The two homeodomains of the ZmHox2a gene from maize originated as an internal gene duplication and have evolved different target site specificities.

The maize ZmHox2a gene encodes two homeodomains which originated by a 699 bp duplication within an ancestral precursor. The sequences of the two ZmHox2a homeodomains are highly diverged in the N-terminal arm, while residues in the helical part have mostly been conserved. We show here that both ZmHox2a homeodomains are functional DNA-binding motifs but exhibit different target site specificities. CASTing experiments reveal a TCCT motif recognized by HD1 but a GATC tetranucleotide as the recognition sequence of HD2. Mutation of the central nucleotides in both tetranucleotide core motifs abolishes DNA binding. A domain swap experiment indicates that target site specificity is achieved in a combinatorial manner by the contributions of the diverged N-terminal arms together with the slightly different recognition helices. Computer modelling suggests that K47 and H54 in the recognition helices preferentially contact the bases at the 3'-terminus of the tetranucleotide target sequences.

Crystal structure of PHO4 bHLH domain-DNA complex: flanking base recognition.

The crystal structure of a DNA-binding domain of PHO4 complexed with DNA at 2.8 A resolution revealed that the domain folds into a basic-helix-loop-helix (bHLH) motif with a long but compact loop that contains a short alpha-helical segment. This helical structure positions a tryptophan residue into an aromatic cluster so as to make the loop compact. PHO4 binds to DNA as a homodimer with direct reading of both the core E-box sequence CACGTG and its 3'-flanking bases. The 3'-flanking bases GG are recognized by Arg2 and His5. The residues involved in the E-box recognition are His5, Glu9 and Arg13, as already reported for bHLH/Zip proteins MAX and USF, and are different from those recognized by bHLH proteins MyoD and E47, although PHO4 is a bHLH protein.

HERITABILITY OF BLUE TIT TARSUS LENGTH FROM NORMAL AND CROSS-FOSTERED BROODS

EvolutionVolume 36, Issue 2 p. 418-419 Note and CommentFree Access HERITABILITY OF BLUE TIT TARSUS LENGTH FROM NORMAL AND CROSS-FOSTERED BROODS André A. Dhondt, André A. Dhondt Department of Biology, Universitaire Instelling Antwerpen, B-2610, Wilrijk, BelgiumSearch for more papers by this author André A. Dhondt, André A. Dhondt Department of Biology, Universitaire Instelling Antwerpen, B-2610, Wilrijk, BelgiumSearch for more papers by this author First published: March 1982 https://doi.org/10.1111/j.1558-5646.1982.tb05061.xCitations: 42 Corresponding Editor: D. B. Wake AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume36, Issue2March 1982Pages 418-419 ReferencesRelatedInformation

A Low Inductance 120 KV Energy Storage Module

A low inductance 120 kV energy storage module has been developed at the Naval Research Laboratory. Fifty of these modules operating in parallel will form the 300 kJ capacitor bank for the fast reversal field of the SWAP experiment. The module comprises two 60 kV, 1.85 ?F capacitors operating as a modified Marx-type circuit. The self-inductance of the module, including the output switch and cable terminations, is approximately 110 nH. This arrangement has the advantage of using 60 kV power supplies, charging circuit and capacitors for a 120 kV capacitor bank and eliminates one of the series fast synchronized switches with a concomitant inductance reduction. The bank construction cost has been drastically reduced by the module physical arrangement. The second stage capacitor is mounted upside down directly on the first stage capacitor. The first stage capacitor therefore acts as both the support and insulator for the second stage capacitor whose case rises to 120 kV above ground at discharge.