Sexual differentiation in higher plants

As in all organogenetic studies, sex differentiation implies three fields of research: the identification of macsomalecular markers specific for stamens or carpels }expression program); the analysis of the signals (such as phytohormones{ inducing them: and finally the knowledge of the regulator genes (sex detsrmination genes).
In dioecious plants, sex determination (male and female genes or combinations of heterochromosomes) occurs at the fertilization stage. These regulators probably act early by means of inducers; their action is perceptible mainly when reproductive organs develop, and the existence of target cells able to respond to the inductive message is obvious. Experimental control of sex expression by phytohormones leads to the repression of the normal organogenetic program (induced by the presence of the sex genes) arrd to the induction of the opposite program (normally incompatible with these genes).
Analogous mechanisms occur in monoecious plants, but here the male program is always expressed before the female one. Experimental control of sex expression suggests that the successive induction of first male and then the female program results from inverse gradients of male and female signals. Sex differentiation studies of monaeciousness are more difficult than studies on dioeciousness since, in all the cells. not only male and female programs co-exist as in dioeciousness, but also male and female sex penes and signals.
The present model has been tested in the dioecious Mercurialisannua (2n = 16) and some other species, and the results that have been established are described.     

WNK kinases sense molecular crowding and rescue cell volume via phase separation

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Macromolecular rate theory (MMRT) provides a thermodynamics rationale to underpin the convergent temperature response in plant leaf respiration

Temperature is a crucial factor in determining the rates of ecosystem processes, for example, leaf respiration (R) – the flux of plant respired CO₂ from leaves to the atmosphere. Generally, R increases exponentially with temperature and formulations such as the Arrhenius equation are widely used in earth system models. However, experimental observations have shown a consequential and consistent departure from an exponential increase in R. What are the principles that underlie these observed patterns? Here, we demonstrate that macromolecular rate theory (MMRT), based on transition state theory (TST) for enzyme‐catalyzed kinetics, provides a thermodynamic explanation for the observed departure and the convergent temperature response of R using a global database. Three meaningful parameters emerge from MMRT analysis: the temperature at which the rate of respiration would theoretically reach a maximum (the optimum temperature, T_opt), the temperature at which the respiration rate is most sensitive to changes in temperature (the inflection temperature, T_inf) and the overall curvature of the log(rate) versus temperature plot (the change in heat capacity for the system, ). On average, the highest potential enzyme‐catalyzed rates of respiratory enzymes for R are predicted to occur at 67.0 ± 1.2°C and the maximum temperature sensitivity at 41.4 ± 0.7°C from MMRT. The average curvature (average negative ) was ?1.2 ± 0.1 kJ mol^?1 K^?1. Interestingly, T_opt, T_inf and appear insignificantly different across biomes and plant functional types, suggesting that thermal response of respiratory enzymes in leaves could be conserved. The derived parameters from MMRT can serve as thermal traits for plant leaves that represent the collective temperature response of metabolic respiratory enzymes and could be useful to understand regulations of R under a warmer climate. MMRT extends the classic TST to enzyme‐catalyzed reactions and provides an accurate and mechanistic model for the short‐term temperature response of R around the globe.     

Macromolecular domains containing nuclear protein p107 and U-snRNP protein p28: Further evidence for an in situ nuclear matrix

Summary Polyclonal antibodies have been produced which react with a nuclear protein having a molecular weight of 107kD and a pl of 8.7–8.8 (designated p107). This protein is shown to be a component of the residual ribonucleoprotein (RNP) network of the nuclear matrix. P107 localized exclusively to the nuclear interior but not within nucleolar or chromatin domains. We have taken advantage of this unique probe to examine whether the RNP network of the isolated nuclear matrix has a physical counterpart in situ. We show that RNA, p107, divalent cations and the 28 kD Sm antigen of U-snRNPs are components of in situ macromolecular assemblies. While the morphology and intranuclear distribution of these assemblies are insensitive to the removal of chromatin, they are markedly altered by degradation of RNA. Digestion in situ of RNA in the presence of EDTA followed by extraction with high ionic strength buffers solubilized the components of these assemblies. Electron microscopic and immunobiochemical data are presented which support the concept that the residual RNP network of the nuclear matrix is an isolate of a pre-existing structure, and that perturbations in this internal network can be created by RNA degradation, depletion of essential metal ions and proteolysis.Abbreviations CRLM polyclonal chicken antibody raised against rat liver nuclear matrix - Sm monoclonal antibody specific for the 28 kd protein antigen of U1, U2, U4, U5 and U6 snRNPs - hnRNP ribonucleoprotein particles containing hnRNA - snRNP ribonucleoprotein particles containing snRNA - PBS phosphate buffered saline - PMSF phenylmethylsulfonyl fluoride - PAGE polyacrylamide gel electrophoresis - EDTA ethylenediaminetetraacetic acid - VRC vanadium ribonucleoside complex - BSA bovine serum albumin - DMSO dimethylsulfoxide - HS high salt buffer - LS low salt buffer     

Growth ofMycobacterium smegmatis in minimal and complete media

The growth patterns ofMycobacterium smegmatis SN2 in a minimal medium and in nutrient broth have been compared. The growth was monitored by absorbancy (Klett readings), colony forming units, wet weight and content of DNA, RNA and protein. During the early part of the growth cycle, the bacteria had higher wet weight and macromolecular content in nutrient broth than in minimal media. During the latter half of the growth cycle however, biosynthesis stopped much earlier in nutrient broth and the bacteria had a much lower content of macromolecules than in the minimal medium. In both the media, a general pattern of completing biosynthesis rapidly in the initial phase and a certain amount of cell division at a later time involving the distribution of preformed macromolecules was seen. The possible adaptive significance of this observation has been discussed.     

Stoichiometry of Nucleotide Binding to Proteasome AAA+ ATPase Hexamer Established by Native Mass Spectrometry

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Highlights

• •All six binding sites in PAN^WT are occupied by ADP- or ATP-type nucleotides.
• •PAN^KA Walker A mutant substoichiometrically binds ATP- but not ADP-type nucleotides.
• •PAN hexamer dissociation of the solution origin characteristics was observed in MS.
• •We posit that the PAN hexamer dissociation proceeds within the ESI droplets.

     

Enhanced validation of small-molecule ligands and carbohydrates in the Protein Data Bank

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Bayesian Weighing of Electron Cryo-Microscopy Data for Integrative Structural Modeling

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Assessing the reconstruction of macromolecular assemblies with toleranced models

We introduce toleranced models (TOMs), a generic and versatile framework meant to handle models of macromolecular assemblies featuring uncertainties on the shapes and the positions of proteins. A TOM being a continuum of nested shapes, the inner (resp. outer) ones representing high (low) confidence regions, we present topological and geometric statistics assessing features of this continuum at multiple scales. While the topological statistics qualify contacts between instances of protein types and complexes involving prescribed protein types, the geometric statistics scale the geometric accuracy of these complexes. We validate the TOM framework on recent average models of the entire nuclear pore complex (NPC) obtained from reconstruction by data integration, and confront our quantitative analysis against experimental findings related to complexes of the NPC, namely the Y-complex, the T-complex, and the Nsp1-Nup82-Nup159 complex. In the three cases, our analysis bridges the gap between global qualitative models of the entire NPC, and atomic resolution models or putative models of the aforementioned complexes. In a broader perspective, the quantitative assessments provided by the TOM framework should prove instrumental to implement a virtuous loop "model reconstruction-model selection", in the context of reconstruction by data integration.     

Antimicrobial Actions of Human and Macaque Sperm Associated Antigen (SPAG) 11 Isoforms: influence of the N-terminal peptide

In addition to their role in sperm maturation, recent evidence has indicated that epididymal proteins have a role in male reproductive tract innate immunity. Herein we demonstrate that human and macaque epididymal protein isoforms in the SPAG (sperm associated antigen) 11 family, full length SPAG11C, K and L exhibit potent antibacterial activity against E. coli. Analysis of activities of the N- and C-terminal domains revealed that the human N-terminal peptide is bactericidal, while the C-terminal domains that contain the defensin-like 6 cysteine array in SPAG11C and partial arrays in SPAG11K and SPAG11L, lack antibacterial activity. The N-terminal peptide does not appear to contain all the determinants of activity since full-length human SPAG11C is more active than the isolated N-terminal peptide and since sulfhydryl reduction and alkylation, which would affect primarily the C-terminal peptides, completely abolished activities of the whole proteins. These results suggest that the structure conferred by the disulfide bonds in human SPAG11C contributes to the antibacterial activity of the whole molecule. The activities of the N-terminal peptide and of full length human SPAG11C were somewhat reduced in increasing NaCl concentrations. In contrast, the antibacterial activities of full length macaque SPAG11C, K and L were unaffected by the presence of NaCl suggesting a mechanism in the macaque that is less dependent upon electrostatic interactions. SPAG11C, K and L disrupted E. coli membranes but had no effect on erythrocyte membranes. Inhibition of E. coli RNA, DNA and protein synthesis by nonlethal concentrations of SPAG11 isoforms indicated an additional mechanism of bacterial killing. Abbreviation: SPAG11, sperm associated antigen 11; CFUs, colony forming units; NPN, N-phenyl-1-napthylamine; diSC3-5, 3,5-dipropylthiadicarbocyanine iodide; IAA, iodoacetamide; BME, β-mercaptoethanol