Reversible inhibition of translation by Xenopus oocyte-specific proteins

A characteristic of growing oocytes of all animal species is the synthesis and accumulation of messenger RNA which is destined to be used primarily by the early embryo. The mechanism(s) which regulates the translation of this maternal mRNA remains unknown. However, the inability of the oocyte to translate all of its putative mRNA has been attributed to at least three limitations: (1) The rate of translation is limited by the availability of components of the translational apparatus other than mRNA, (2) the structural organization of the mRNA prevents translation, and (3) proteins associated with the mRNA prevent translation. Several investigators have suggested that proteins associated with maternal mRNA suppress translation in sea urchin eggs, although others claim that such results may be due to experimental artefacts. Oocyte-specific proteins have been identified in association with non-translating poly(A)+ mRNAs from Xenopus laevis oocytes, and we report here that when these proteins are reconstituted with mRNAs in vitro the translation of the mRNAs in vitro is reversibly repressed. The implication is that these proteins are involved in the regulation of translation of stored maternal mRNAs.     

低氧胁迫对光棘球海胆Mesocentrotus nudus致死性及生理机能的影响

全球近海底层水体低氧现象日趋频繁，严重影响到海洋生态系统的稳定性。为掌握光棘球海胆Mesocentrotus nudus对低氧胁迫的耐受能力，通过室内生态模拟实验，分析3种规格的光棘球海胆，即大规格海胆平均体质量(91.41±3.31) g、中规格(60.79±3.42) g和小规格(4.15±0.18) g，在3个溶解氧(Dissolved Oxygen，DO)浓度水平(0.5 mg/L、2.0 mg/L、3.0 mg/L)胁迫下的行为、存活、半致死时间(Median Lethal Time，LT₅₀)以及超氧化物歧化酶(Superoxide Dismutase，SOD)活力。结果表明，在DO 0.5 mg/L和DO 2.0 mg/L低氧胁迫下，海胆行为表现异常，低氧初期海胆表现为管足活力下降、吸附力降低，移动能力变弱，同时伴随棘脱落，随着低氧暴露时间延长，海胆各项活动能力进一步减弱，直至死亡；在DO 3.0 mg/L胁迫下，中、大规格海胆无死亡现象。3种规格海胆死亡率随低氧暴露时间延长都有所增加，其中在DO 0.5 mg/L胁迫下，3种规格海胆的最终死亡率都达到100%，但在DO 2.0 mg/L胁迫下，随低氧胁迫时间的延长，3种规格海胆都对低氧胁迫表现出不同程度的适应，很少死亡。3种规格海胆的LT₅₀在DO 0.5 mg/L和DO 2.0 mg/L两种低氧胁迫水平下存在差异，DO 0.5 mg/L胁迫下，大规格海胆的LT₅₀为70.50 h，中规格为34.64 h，小规格为34.02 h；DO 2.0 mg/L胁迫下，中规格海胆的LT₅₀延长至120.88 h，小规格海胆延长至107.15 h。此外，中规格海胆在两种低氧胁迫水平下，体内的超氧化物歧化酶活力均发生了明显变化，说明低氧胁迫已影响到光棘球海胆的抗氧化系统。本研究初次掌握了光棘球海胆在不同环境低氧胁迫下的生存和低氧适应情况，可为低氧海域光棘球海胆种群维持以及海胆养殖业的健康发展提供参考。     

A gene regulatory network controlling the embryonic specification of endoderm

Specification of endoderm is the prerequisite for gut formation in the embryogenesis of bilaterian organisms. Modern lineage labelling studies have shown that in the sea urchin embryo model system, descendants of the veg1 and veg2 cell lineages produce the endoderm, and that the veg2 lineage also gives rise to mesodermal cell types. It is known that Wnt/β-catenin signalling is required for endoderm specification and Delta/Notch signalling is required for mesoderm specification. Some direct cis-regulatory targets of these signals have been found and various phenomenological patterns of gene expression have been observed in the pre-gastrular endomesoderm. However, no comprehensive, causal explanation of endoderm specification has been conceived for sea urchins, nor for any other deuterostome. Here we propose a model, on the basis of the underlying genomic control system, that provides such an explanation, built at several levels of biological organization. The hardwired core of the control system consists of the cis-regulatory apparatus of endodermal regulatory genes, which determine the relationship between the inputs to which these genes are exposed and their outputs. The architecture of the network circuitry controlling the dynamic process of endoderm specification then explains, at the system level, a sequence of developmental logic operations, which generate the biological process. The control system initiates non-interacting endodermal and mesodermal gene regulatory networks in veg2-derived cells and extinguishes the endodermal gene regulatory network in mesodermal precursors. It also generates a cross-regulatory network that specifies future anterior endoderm in veg2 descendants and institutes a distinct network specifying posterior endoderm in veg1-derived cells. The network model provides an explanatory framework that relates endoderm specification to the genomic regulatory code.     

海胆早期神经发育及有机磷农药对其神经毒性作用的研究进展

 早期发育阶段是有机磷农药神经毒性作用最敏感的时期,海胆的胚胎和幼虫为研究有机磷农药对早期发育阶段的神经毒性作用提供了一种理想的模型。本文介绍了海胆的早期神经发育过程,综述了神经系统对海胆早期发育的调控作用,结合近年来国内外的研究,阐述了有机磷农药对海胆早期发育的影响及其神经毒性作用机制,并展望了该领域的研究方向。     

Identification of diploid endosperm in an early angiosperm lineage

In flowering plants, the developmental and genetic basis for the establishment of an embryo-nourishing tissue differs from all other lineages of seed plants. Among extant nonflowering seed plants (conifers, cycads, Ginkgo, Gnetales), a maternally derived haploid tissue (female gametophyte) is responsible for the acquisition of nutrients from the maternal diploid plant, and the ultimate provisioning of the embryo. In flowering plants, a second fertilization event, contemporaneous with the fusion of sperm and egg to yield a zygote, initiates a genetically biparental and typically triploid embryo-nourishing tissue called endosperm. For over a century, triploid biparental endosperm has been viewed as the ancestral condition in extant flowering plants. Here we report diploid biparental endosperm in Nuphar polysepalum, a basal angiosperm. We show that diploid endosperms are common among early angiosperm lineages and may represent the ancestral condition among flowering plants. If diploid endosperm is plesiomorphic, the triploid endosperms of the vast majority of flowering plants must have evolved from a diploid condition through the developmental modification of the unique fertilization process that initiates endosperm.     

Production of transgenic rabbits, sheep and pigs by microinjection

Direct microinjection has been used to introduce foreign DNA into a number of terminally differentiated cell types as well as embryos of several species including sea urchin, Candida elegans, Xenopus, Drosophila and mice. Various genes have been successfully introduced into mice including constructs consisting of the mouse metallothionein-I (MT) promoter/regulator region fused to either the rat or human growth hormone (hGH) structural genes. Transgenic mice harbouring such genes commonly exhibit high, metal-inducible levels of the fusion messenger RNA in several organs, substantial quantities of the foreign growth hormone in serum and enhanced growth. In addition, the gene is stably incorporated into the germ line, making the phenotype heritable. Because of the scientific importance and potential economic value of transgenic livestock containing foreign genes, we initiated studies on large animals by microinjecting the fusion gene, MT-hGH, into the pronuclei or nuclei of eggs from superovulated rabbits, sheep and pigs. We report here integration of the gene in all three species and expression of the gene in transgenic rabbits and pigs.     

Duplication and remoulding of tRNA genes during the evolutionary rearrangement of mitochondrial genomes

During the evolution of sea urchins, a transfer RNA gene lost its tRNA function and became part of a protein-coding gene. This functional loss of a tRNA with specificity for one group of leucine codons (CUN, where N is any base) was accompanied by the gain of a new tRNA with that specificity. The new tRNA gene for CUN codons appears to have evolved by duplication and divergence from a tRNA gene specific for another group of leucine codons (UUR, where R is a purine). These proposals account for (1) the strong sequence resemblance between the modern tRNA genes for CUN and UUR codons in Paracentrotus, (2) the altered location of the CUN gene in mitochondrial DNA of this urchin, and (3) the persistence of a 72-base pair sequence containing a trace of the old CUN gene at its original location. The old CUN gene now codes for an extra 24 amino acids at the amino end of subunit 5 in NADH dehydrogenase. Besides giving clues about the mechanisms by which tRNA genes move during mitochondrial DNA evolution, this finding leads us to propose a pathway relating the arrangements of other genes in mitochondrial DNAs from four animal phyla.     

Positive selection of a gene family during the emergence of humans and African apes

Gene duplication followed by adaptive evolution is one of the primary forces for the emergence of new gene function. Here we describe the recent proliferation, transposition and selection of a 20-kilobase (kb) duplicated segment throughout 15 Mb of the short arm of human chromosome 16. The dispersal of this segment was accompanied by considerable variation in chromosomal-map location and copy number among hominoid species. In humans, we identified a gene family (morpheus) within the duplicated segment. Comparison of putative protein-encoding exons revealed the most extreme case of positive selection among hominoids. The major episode of enhanced amino-acid replacement occurred after the separation of human and great-ape lineages from the orangutan. Positive selection continued to alter amino-acid composition after the divergence of human and chimpanzee lineages. The rapidity and bias for amino-acid-altering nucleotide changes suggest adaptive evolution of the morpheus gene family during the emergence of humans and African apes. Moreover, some genes emerge and evolve very rapidly, generating copies that bear little similarity to their ancestral precursors. Consequently, a small fraction of human genes may not possess discernible orthologues within the genomes of model organisms.     

Stabilization of sea urchin flagellar microtubules by histone H1.

Complex microtubule assemblies are essential components of eukaryotic cilia and flagella. They are extremely stable and are not affected by agents that normally induce polymer disassembly. The molecular basis of this microtubular stability is unknown, and it is not related to any feature of the constitutive tubulin. In sea urchin sperm flagella, axonemal microtubules are found to be stabilized by a protein identical to histone H1, a result that defines a new role for this histone and provides evidence for a concerted evolution of chromatin and microtubular structures.     

Genetic evidence for female host-specific races of the common cuckoo

The common cuckoo Cuculus canorus is divided into host-specific races (gentes). Females of each race lay a distinctive egg type that tends to match the host's eggs, for instance, brown and spotted for meadow pipit hosts or plain blue for redstart hosts. The puzzle is how these gentes remain distinct. Here, we provide genetic evidence that gentes are restricted to female lineages, with cross mating by males maintaining the common cuckoo genetically as one species. We show that there is differentiation between gentes in maternally inherited mitochondrial DNA, but not in microsatellite loci of nuclear DNA. This supports recent behavioural evidence that female, but not male, common cuckoos specialize on a particular host, and is consistent with the possibility that genes affecting cuckoo egg type are located on the female-specific W sex chromosome. Our results also support the ideas that common cuckoos often switched hosts during evolution, and that some gentes may have multiple, independent origins, due to colonization by separate ancestral lineages.     

Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle

To understand the molecular basis of microtubule-associated motility during mitosis, the mechanochemical factors that generate the relevant motile force must be identified. Myosin, the ATPase that interacts with actin to produce the force for muscle contraction and other forms of cell motility, is believed to be involved in cytokinesis but not in mitosis. Dynein, the mechanochemical enzyme that drives microtubule sliding in eukaryotic cilia and flagella, has been identified in the cytoplasm of sea urchin eggs, but the evidence that it is involved in cytoplasmic microtubule-based motility (rather than serving as a precursor for embryonic cilia) is equivocal. Microtubule-associated ATPases have been prepared from other tissues, but their role in cytoplasmic motility is also unknown. Recent work on axoplasmic transport, however, has led to the identification of a novel mechanochemical protein called kinesin, which is thought to generate the force for moving vesicles along axonal microtubules. These results suggest that kinesin may also be a mechanochemical factor for non-axoplasmic forms of microtubule-based motility, such as mitosis. We describe here the identification and isolation of a kinesin-like protein from the cytoplasm of sea urchin eggs. We present evidence that this protein is localized in the mitotic spindle, and propose that it may be a mechanochemical factor for some form of motility associated with the mitotic spindle.     

Compensatory mutations suggest that base-pairing with a small nuclear RNA is required to form the 3' end of H3 messenger RNA

Processing of the 3' end of sea urchin H3 histone pre-mRNA requires conserved sequence elements and the presence of U7 snRNA. A mutation in the conserved CAAGAAGA sequence of the H3 pre-mRNA that renders 3' processing of this precursor defective is shown to be suppressed by a compensatory change in the U7 snRNA, restoring the base-pairing potential of the two RNAs. RNA-RNA contacts between these two molecules appear to be an essential feature of the 3' processing reaction.     

Gene transfer to the nucleus and the evolution of chloroplasts

Photosynthetic eukaryotes, particularly unicellular forms, possess a fossil record that is either wrought with gaps or difficult to interpret, or both. Attempts to reconstruct their evolution have focused on plastid phylogeny, but were limited by the amount and type of phylogenetic information contained within single genes. Among the 210 different protein-coding genes contained in the completely sequenced chloroplast genomes from a glaucocystophyte, a rhodophyte, a diatom, a euglenophyte and five land plants, we have now identified the set of 45 common to each and to a cyanobacterial outgroup genome. Phylogenetic inference with an alignment of 11,039 amino-acid positions per genome indicates that this information is sufficient--but just rarely so--to identify the rooted nine-taxon topology. We mapped the process of gene loss from chloroplast genomes across the inferred tree and found that, surprisingly, independent parallel gene losses in multiple lineages outnumber phylogenetically unique losses by more that 4:1. We identified homologues of 44 different plastid-encoded proteins as functional nuclear genes of chloroplast origin, providing evidence for endosymbiotic gene transfer to the nucleus in plants.     

真核生物中支架蛋白家族(IQGAP family)的分子进化分析

支架蛋白家族(IQGAP family)广泛存在于真核生物中,在细胞的信号转导、骨架运动、细胞分裂的过程中都有着重要功能.研究分析IQGAP蛋白家族的分子进化,有助于深入全面地了解整个IQGAP蛋白家族在不同物种中功能.通过使用相关数据库,在35个真核生物物种中检索到了70个支架蛋白家族成员的序列信息,并对其进行了系统的分子进化研究.我们发现:支架蛋白家族基因广泛存在于后生动物(Metazoan)、真菌(Fungi)、变形虫界(Amoebozoa)以及其他一些真核生物中,提示支架蛋白存在于最后的真核生物的共同祖先(Last Eukaryotic Common Ancestor,LECA).系统发育分析表明脊椎动物的IQGAP蛋白和非脊椎动物中代表性IQGAP基因共享一个共同的祖先基因,单个的IQGAP祖先基因在早期脊椎动物中(在四足总纲和硬骨鱼纲的分离之前),发生了两次基因复制事件,导致现存的脊椎动物中3组IQGAP基因(IQGAP1,IQGAP2,IQGAP3)产生.     

Correlation between the ATPase and microtubule translocating activities of sea urchin egg kinesin

Coupling between ATP hydrolysis and microtubule movement was demonstrated several years ago in flagellar axonemes and subsequent studies suggest that the relevant microtubule motor, dynein, uses ATP to drive microtubule sliding by a cross-bridge mechanism analogous to that of myosin in muscles. Kinesin, a microtubule-based motility protein which may participate in organelle transport and mitosis, binds microtubules in a nucleotide-sensitive manner, and requires hydrolysable nucleotides to translocate microtubules over a glass surface. Recently, neuronal kinesin was shown to possess microtubule-activated ATPase activity although coupling between ATP hydrolysis and motility was not demonstrated. Here we report that sea urchin egg kinesin, prepared either with or without a 5'-adenylyl imidodiphosphate(AMPPNP)-induced microtubule binding step, also possesses significant microtubule-activated ATPase activity when Mg-ATP is used as a substrate. This ATPase activity is inhibited in a dose-dependent manner by addition of Mg-free ATP, by chelation of Mg2+ with EDTA, by addition of Na3VO4, or by addition of AMPPNP with or without Mg2+. Addition of these same reagents also inhibits the microtubule-translocating activities of sea urchin egg kinesin in a dose-dependent manner, supporting the hypothesis that kinesin-driven motility is coupled to the microtubule-activated Mg2+-ATPase activity.     

Regions of variant histone His2AvD required for Drosophila development.

One way in which a distinct chromosomal domain could be established to carry out a specialized function is by the localized incorporation of specific histone variants into nucleosomes. H2AZ, one such variant of the histone protein H2A, is required for the survival of Drosophila melanogaster, Tetrahymena thermophila and mice (R. Faast et al., in preparation). To search for the unique features of Drosophila H2AZ (His2AvD, also referred to as H2AvD) that are required for its essential function, we have performed amino-acid swap experiments in which residues unique to Drosophila His2AvD were replaced with equivalently positioned Drosophila H2A.1 residues. Mutated His2AvD genes encoding modified versions of this histone were transformed into Drosophila and tested for their ability to rescue null-mutant lethality. We show that the unique feature of His2AvD does not reside in its histone fold but in its carboxy-terminal domain. This C-terminal region maps to a short alpha-helix in H2A that is buried deep inside the nucleosome core.     

Translocation of a localized maternal mRNA to the vegetal pole of Xenopus oocytes

A prominent hypothesis in embryology is that localized maternal factors are important in specifying cell fate. There are, however, only a few examples of maternal molecules that have been shown to be localized and very little is known about how such factors are physically localized within an egg (for review see ref. 1). Previously, cDNA clones were obtained for a class of localized maternal mRNAs from Xenopus laevis. These mRNAs are unusual in that they are concentrated at either the animal or vegetal pole of unfertilized eggs. In the present study the synthesis and intracellular distribution of one of them, Vg1, has been examined during oogenesis. The results show that Vg1 mRNA is localized as a crescent at the vegetal pole of mature oocytes. Surprisingly, this mRNA is uniformly distributed in the cytoplasm of immature oocytes. These findings suggest that a single cell, the frog oocyte, has some mechanism for translocating specific RNAs like Vg1. The process that moves Vg1 mRNA is evidently a cytoplasmic localization machinery which is not directly coupled to the synthesis of Vg1 RNA.