Selectivity of yolk protein uptake: Comparison of vitellogenins of two insects

The vitellogenins of Hyalophora cecropia and Blattella germanica have similar Stokes' radii (69 and 75Å), sedimentation coefficients (15.9 and 16.8S) and isoelectric points (pH 5.7 and 5.0), and share similar amino acid compositions with vitellogenins of other animals. The two vitellogenins show no immunological crossreaction. Blattella oöcytes take up their own vitellogenin in vivo at a rapid rate and that of Hyalophora at a low rate comparable to that of a non-vitellogenic protein. Hyalophora oöcytes take up their own vitellogenin rapidly in vitro and Blattella vitellogenin only at a negligible rate. Molecular weights, shapes and charges of the vitellogenins are similar to non-vitellogenins and form no basis for selective uptake.     

Derivation of major yolk proteins from parental vitellogenins and alternative processing during oocyte maturation in Fundulus heteroclitus

Various Coomassie blue-staining yolk proteins (YPs) present in oocytes and eggs of Fundulus heteroclitus, a teleost that produces low hydrated, demersal eggs (benthophil species), were subjected to N-terminal microsequencing. Four YPs were N-terminally blocked, while five yielded sequence information. Of the latter, four corresponded to internal sequences of vitellogenin 1 (Vg1), whereas a fifth band corresponded to the N-terminal sequence of Vg2. Phosphorylated YPs (phosvitins and phosvettes) derived from the polyserine domain of Vg were not successfully sequenced. The major N-terminally blocked 122-and 103-kDa YPs both represented the lipovitellin heavy chain of Vg1 (LvH1), and thus most of the oocyte YPs were derived from Vg1. During oocyte maturation in vivo and in vitro, the LvH1 122 is degraded, concomitant with an increased enzymatic activity of cathepsin B, while the 45-kDa YP is converted to a 42-kDa YP. The LvH1 122 was found to contain a consensus site for proteolytic degradation (PEST) near its C-terminus, which is missing from its stable, but truncated twin sequence, LvH1 103. We suggest that this site becomes exposed to cathepsin B during the hydration process that accompanies oocyte maturation and renders the LvH1 122 susceptible to proteolysis. PEST sites are found in Vg sequences from other benthophil fish, whereas, interestingly, they are missing in marine teleosts that spawn highly hydrated, pelagic eggs (pelagophil species), displaying a different pattern of Vg incorporation into YPs and LvH1 and LvH2 processing to that found in F. heteroclitus. Thus, different models of Vg/YP precursor/product relationship and further processing during oocyte maturation and hydration are proposed for pelagophil and benthophil teleosts.     

Multiple vitellogenins (Vgs) in mosquitofish (Gambusia affinis): identification and characterization of three functional Vg genes and their circulating and yolk protein products

The objectives of this study were to characterize multiple forms of vitellogenin (Vg) in mosquitofish (Gambusia affinis) and to discover the fate of each Vg during its processing into product yolk proteins. Two Vg preparations, with apparent masses of 600 kDa (600 Vg) and 400 kDa (400 Vg), were isolated from the plasma of fish treated with estradiol-17beta (E(2)) by various chromatographic procedures. Immunological analyses verified the presence of two different Vg proteins (600 VgA and 600 VgB) in the 600 Vg preparation and of a single protein in the 400 Vg preparation. Three major yolk proteins (Yps) with apparent masses of 560, 400, and 28 kDa were observed in extracts of ovarian follicles from vitellogenic females. Immunological analyses demonstrated that the 400 Vg underwent no change in native mass after being incorporated into oocytes. The 600 Vgs gave rise to a 28 kDa beta'-component and a native 560 kDa Yp, which was heterodimeric in structure, consisting of two types of complexes between phosvitin (Pv) and lipovitellin (Lv) heavy- and light-chains. Full-length cDNAs encoding the 600 VgA, 600 VgB, and 400 Vg were isolated from a liver cDNA library of E(2) treated fish. Similar to the zebrafish vg3 gene, the 400 Vg cDNA lacked a Pv domain and was classified as an incomplete or phosvitinless (C-type) Vg. The deduced primary structures of 600 VgA and 600 VgB were complete, and these were categorized as type A and type B Vgs, respectively, according to our recent classification scheme. This is the first report on the characterization of three functional Vg genes and their circulating and yolk protein products in any vertebrate species.     

Structural and functional analysis of HtrA1 and its subdomains

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Highlights? IGFBP-like domain structure suggests that it is incompetent for IGF binding ? Kazal-like domain structure shows distorted “P1” loop incompetent for inhibition ? Protease domain shows catalytic readiness in apo form ? SAXS envelope for full-length HtrA1     

Cardiolipin binding in bacterial respiratory complexes: Structural and functional implications

The structural and functional integrity of biological membranes is vital to life. The interplay of lipids and membrane proteins is crucial for numerous fundamental processes ranging from respiration, photosynthesis, signal transduction, solute transport to motility. Evidence is accumulating that specific lipids play important roles in membrane proteins, but how specific lipids interact with and enable membrane proteins to achieve their full functionality remains unclear. X-ray structures of membrane proteins have revealed tight and specific binding of lipids. For instance, cardiolipin, an anionic phospholipid, has been found to be associated to a number of eukaryotic and prokaryotic respiratory complexes. Moreover, polar and septal accumulation of cardiolipin in a number of prokaryotes may ensure proper spatial segregation and/or activity of proteins. In this review, we describe current knowledge of the functions associated with cardiolipin binding to respiratory complexes in prokaryotes as a frame to discuss how specific lipid binding may tune their reactivity towards quinone and participate to supercomplex formation of both aerobic and anaerobic respiratory chains. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).     

燕辽生物群脊椎动物的多样性及与其他生物群的对比分析*

燕辽生物群已发现脊椎动物54属58种, 包括鱼类、两栖类、爬行类、哺乳类等, 但其脊椎动物多样性及其成因机制还未有详细研究。本文对该生物群脊椎动物进行统计分析, 并与同时代的其他生物群脊椎动物类型进行对比, 这为认识燕辽生物群脊椎动物的多样性及其成因提供了重要的证据。早期代表道虎沟生物群与晚期代表玲珑塔生物群虽存在时代上的传承关系, 但生物组合特征明显不同。对比燕辽生物群与相近时代的新疆五彩湾动物群和四川大山铺恐龙动物群, 脊椎动物组合特征差异显著。燕辽生物群恐龙类群主要以小型兽脚类恐龙为主, 还包括一些小型鸟臀类恐龙。另外还具有非常丰富的翼龙和哺乳动物。脊椎动物生态多样性高, 适应飞行、树栖、水生、穴居等多种生活方式, 但是脊椎动物的类型与同时代的相近地区明显不同。翼龙、恐龙和哺乳动物等类群都展现出独特的生物组合特征。有证据表明该时期东亚地区与其他地区可能存在一定程度的地理隔离, 结合陆生脊椎动物组合特征推测燕辽生物群脊椎动物与外界可能存在一定的交流障碍。     

Vertebrate and nematode genes coding for yolk proteins are derived from a common ancestor

One of the most obvious characteristics of the egg cells of oviparous animals is their large size resulting to a major extent from the deposition of nutritional reserves, mainly constituted of yolk proteins. In general, these are derived from a precursor called vitellogenin, which undergoes posttranslational modifications during secretion and during transport into and storage within the oocytes. Comparative analysis of the structural organization of the vitellogenin gene and of its product in different species shows that the vitellogenin gene is very ancient and that in vertebrates the gene may have more resemblance to the earliest gene than in invertebrates.     

Myosin V attachment to cargo requires the tight association of two functional subdomains

The myosin V carboxyl-terminal globular tail domain is essential for the attachment of myosin V to all known cargoes. Previously, the globular tail was viewed as a single, functional entity. Here, we show that the globular tail of the yeast myosin Va homologue, Myo2p, contains two structural subdomains that have distinct functions, namely, vacuole-specific and secretory vesicle-specific movement. Biochemical and genetic analyses demonstrate that subdomain I tightly associates with subdomain II, and that the interaction does not require additional proteins. Importantly, although neither subdomain alone is functional, simultaneous expression of the separate subdomains produces a functional complex in vivo. Our results suggest a model whereby intramolecular interactions between the globular tail subdomains help to coordinate the transport of multiple distinct cargoes by myosin V.     

Identification of specific functional subdomains within the linker histone H10 C-terminal domain

Linker histone binding to nucleosomal arrays in vitro causes linker DNA to form an apposed stem motif, stabilizes extensively folded secondary chromatin structures, and promotes self-association of individual nucleosomal arrays into oligomeric tertiary chromatin structures. To determine the involvement of the linker histone C-terminal domain (CTD) in each of these functions, and to test the hypothesis that the functions of this highly basic domain are mediated by neutralization of linker DNA negative charge, four truncation mutants were created that incrementally removed stretches of 24 amino acids beginning at the extreme C terminus of the mouse H1(0) linker histone. Native and truncated H1(0) proteins were assembled onto biochemically defined nucleosomal arrays and characterized in the absence and presence of salts to probe primary, secondary, and tertiary chromatin structure. Results indicate that the ability of H1(0) to alter linker DNA conformation and stabilize condensed chromatin structures is localized to specific C-terminal subdomains, rather than being equally distributed throughout the entire CTD. We propose that the functions of the linker histone CTD in chromatin are linked to the characteristic intrinsic disorder of this domain.     

Solution structure of microtubule-associated protein light chain 3 and identification of its functional subdomains

Microtubule-associated protein (MAP) light chain 3 (LC3) is a human homologue of yeast Apg8/Aut7/Cvt5 (Atg8), which is essential for autophagy. MAP-LC3 is cleaved by a cysteine protease to produce LC3-I, which is located in cytosolic fraction. LC3-I, in turn, is converted to LC3-II through the actions of E1- and E2-like enzymes. LC3-II is covalently attached to phosphatidylethanolamine on its C terminus, and it binds tightly to autophagosome membranes. We determined the solution structure of LC3-I and found that it is divided into N- and C-terminal subdomains. Additional analysis using a photochemically induced dynamic nuclear polarization technique also showed that the N-terminal subdomain of LC3-I makes contact with the surface of the C-terminal subdomain and that LC3-I adopts a single compact conformation in solution. Moreover, the addition of dodecylphosphocholine into the LC3-I solution induced chemical shift perturbations primarily in the C-terminal subdomain, which implies that the two subdomains have different sensitivities to dodecylphosphocholine micelles. On the other hand, deletion of the N-terminal subdomain abolished binding of tubulin and microtubules. Thus, we showed that two subdomains of the LC3-I structure have distinct functions, suggesting that MAP-LC3 can act as an adaptor protein between microtubules and autophagosomes.     

Vertebrate browsing impacts in a threatened montane plant community and implications for management

Montane ecosystems are vulnerable to the removal of vegetation cover through browsing by feral or native vertebrate fauna. The highest elevated peaks of the Stirling Range in Western Australia provide habitat for an endemic plant community, Critically Endangered due to plant disease, frequent fire and an emerging threat of browsing by vertebrate fauna. Survey and camera trapping confirmed the herbivorous feral Rabbit (Oryctolagus cuniculus) and native Quokka (Setonix brachyurus) are present. Dietary analysis through faecal examination revealed contrasting diets and implicates native rather than feral species as responsible for impacts on dicotyledonous species, and in particular those of conservation significance. Exclosure experiments conducted over 1 year revealed significant changes in abundance, cover and height of perennial herbs and an increase in growth and/or reproduction of four threatened endemic plants. Detrimental impacts caused by native browsing fauna are not unprecedented and may be attributed to disequilibria in ecosystem processes due to multiple interacting threats. Montane ecosystems may be particularly vulnerable to browsing due to their naturally slow recovery after disturbance and browsing may also create environmental conditions more conducive to plant disease. For plant species with critically low population numbers, the impact of browsing poses a threat to population persistence and undermines investment into other conservation recovery actions. For effective management, it is critical to determine the relative impact of browsing species present. Where native species are implicated, the physical protection of high value assets in wire exclosures is warranted to complement other recovery actions and ensure effective species and community recovery.     

Effect of egg yolk and other reagents upon the zinc of cold-shocked boar spermatozoa

A marked reduction (80.8%) in the zinc uptake by boar spermatozoa cooled to 4 degrees C occurs when the seminal plasma is pretreated with egg yolk-glucose at this temperature. Crude lecithin is less effective (59.8%). Similar pretreatment of the seminal plasma by the polycationic drug Antrypol, which totally removes the zinc-precipitable basic haemagglutinin, does not result in a significant reduction of the sperm zinc uptake at 4 degrees C.