3P045 Analysis of the structural fluctuation in Staphylococcal nuclease and its Δ44-49 mutant: Insight into the enzymatic activity(01B. Protein: Structure & Function,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))
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Antifreeze proteins (AFPs) facilitate cold-survival of the organisms living under freezing environment by inhibiting ice crystal growth.In this study, we focused on a fungal AFP denoted TisAFP secreted from a psychrophilic snow-mold fungus, Typhula ishikariensis.It reported seven isoforms of TisAFP that share a high sequence identity.Significantly, fluorescencebased ice plane affinity (FIPA) analysis showed that the ice binding specificity of TisAFP8 is quite different from the others.To clarify what determines the uniqueness of TisAFP8, we performed mutagenesis experiments on TisAFP8, especially on the residues constructing its putative ice-binding site.The data suggested an importance of inward pointing residues not just outward pointing residues of TisAFP8.Keywords:
Nuclease
We have isolated a novel enhanced-nodulating mutant astray (Ljsym77) from Lotus japonicus. The name astray derives from the non-symbiotic phenotype of this mutant, agravitropic lateral roots that go “astray” against gravity. In this report we evaluated the symbiotic aspects of this mutant in detail. The astray mutant developed approximately twice the number of nodules on a wider area of roots compared with the wild type. Furthermore, the astray mutant demonstrated early initiation of nodule development, which is an unprecedented symbiotic phenotype. The astray seedlings showed normal sensitivity to the general inhibitors of nodulation such as ethylene and nitrate. These results indicate that the astray mutant is distinct from the hypernodulating mutants reported previously, and that the ASTRAY gene acts as an early and negative regulator in the cascade of nodule development.
Lotus japonicus
Nodule (geology)
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To investigate the architecture of the rolled leaf morphology in rice,the features of phenotype and histological level in 2 reverse rolled leaf mutants derived from rice Ac/Ds transposons insertional mutant library were investingated. The results showed that the number of bulliform cells was reduced in 2 reverse rolled leaf mutants(Ad-mutant:adaxial rolled mutant;Ab-mutant:abaxial rolled mutant) compared with the normal flat leaf in wild type,which may be an important cause of the rolled leaf in rice. And the number of parenchyma also was reduced and dehisced to 2 large air cavities in Abmutant. The cellulose content of leaf and culm in 2 rolled leaf mutant and wild type was measured,and the result showed that the cellulose content of leaf and culm in Ab-mutant was significant reduced compared with Ad-mutant and wild type.But the cellulose content of leaf and culm in Ad-mutant was not significantly different from that of wild type. This result suggested that Ab-mutant and Ad-mutant may be different in mutation of gene loci. In addition,the genetic rule of Abmutant and Ad-mutant was analyzed,and the result showed that both of them were controlled by single recessive gene.
Wild type
Genetic Analysis
Parenchyma
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In recent years, the genomes of many different organisms have been fully sequenced and annotated. As a consequence of this information, a number of methods have emerged to study the function of many genes and proteins in parallel. One recent approach for the large-scale analysis of proteins is the use of protein microarrays in which hundreds to thousands of proteins are arrayed and assayed simultaneously. Protein arrays can be used for assessing protein levels and following disease markers, identifying biochemical activities, analyzing post-translational modifications, building interaction networks, and for drug discovery and development. In this review, we discuss the construction of different types of protein arrays, and their numerous and diverse applications.
Protein microarray
Protein Array Analysis
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Proteome
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Protein function prediction
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Guanidine
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(1) Three temperature-sensitive mutants were obtained from a microconidial wild type strain of Neurospora crassa. One of them is irreparable at 34°C; that is, it grows as well as the wild type at 23°C, but does not grow on any (including omplete) medium so far tested at 34°C. This mutant was the major subject of this report.(2) The temperature-sensitive irreparable mutant b39 differs from the parent strain by a single gene mutation. The gene responsible for the irreparable character is located on the first linkage group (mating type chromosome), about 30cM left of the centromere.(3) This mutant does not inhibit the growth of the other temperature-sensitive strains in the heterocaryotic condition.(4) The possibility of the accumulation of inhibitory substances by the mutant is unlikely, because the culture filtrate of the mutant did not inhibit the growth of the mutant at an intermediate temperature.(5) The growth characteristics of the mutant when the culture was transfered from 23°C to 34°C, and from 34°C to 23°C were determined.(6) The nature of the defect in an irreparable mutant was discussed.
Neurospora
Wild type
Crassa
Strain (injury)
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SUMMARY: An investigation was undertaken to determine to what extent the properties of oligosporogenous (Osp) mutants allow them to be considered as a separate class of sporulation mutant, distinct from asporogenous (Sp-) mutants. Of thirty Osp mutants examined, seventeen at least had a phenotype which had previously been identified with a Sp- mutant. The majority of cells in an Osp culture either reached a particular stage in the sporulation process and then stopped, or in some cases went on to produce aberrant forms. Some of these aberrant forms have their counterparts in Sp- mutants described by other authors, but some present new features. The morphological and biochemical sequences were linked so that if the majority of cells were blocked at a certain stage, then the biochemical sequence stopped accordingly. The general similarity in behaviour between the two types of mutant is consistent with the assumption that at least some of the Osp mutants have leaky mutations in genes where mutation can also give rise to Sp- phenotypes. Evidence is presented to suggest that the ability of a cell of an Osp mutant to overcome its block, and so go on to form a spore, is a chance event when that stage in the process is reached. A mutant has been obtained in which the spores are octanol-resistant yet contain no measurable dipicolinate. In several other mutants the spores contained well-developed coat layers, but the cortex was poorly formed or completely missing.
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