The vsp Locus of Mycoplasma bovis: Gene Organization and Structural Features

1999 ◽  
Vol 181 (18) ◽  
pp. 5734-5741 ◽  
Author(s):  
Inessa Lysnyansky ◽  
Konrad Sachse ◽  
Ricardo Rosenbusch ◽  
Sharon Levisohn ◽  
David Yogev
Keyword(s):  
High Frequency ◽  
Sequence Similarity ◽  
Membrane Surface ◽  
Amino Acid Sequences ◽  
High Rate ◽  
Phenotypic Switching ◽  
Upstream Region ◽  
Mycoplasma Bovis ◽  
High Sequence Similarity ◽  
Polypeptide Structure

ABSTRACT Major lipoprotein antigens, known as variable membrane surface lipoproteins (Vsps), on the surface of the bovine pathogenMycoplasma bovis were shown to spontaneously undergo noncoordinate phase variation between ON and OFF expression states. The high rate of Vsp phenotypic switching was also shown to be linked with DNA rearrangements that occur at high frequency in the M. bovis chromosome (I. Lysnyansky, R. Rosengarten, and D. Yogev, J. Bacteriol. 178:5395–5401, 1996). In the present study, 13 single-copyvsp genes organized in a chromosomal cluster were identified and characterized. All vsp genes encode highly conserved N-terminal domains for membrane insertion and lipoprotein processing but divergent mature Vsp proteins. About 80% of eachvsp coding region is composed of reiterated coding sequences that create a periodic polypeptide structure. Eighteen distinct repetitive domains of different lengths and amino acid sequences are distributed within the products of the variousvsp genes that are subject to size variation due to spontaneous insertions or deletions of these periodic units. Some of these repeats were found to be present in only one Vsp family member, whereas other repeats recurred at variable locations in several Vsps. Each vsp gene is also 5′ linked to a highly homologous upstream region composed of two internal cassettes. The findings that rearrangement events are associated with Vsp phenotypic switching and that multiple regions of high sequence similarity are present upstream of the vsp genes and within the vsp coding regions suggest that modulation of the Vsp antigenic repertoire is determined by recombination processes that occur at a high frequency within the vsp locus of M. bovis.

scholarly journals Isolation of Cysteine-Rich Peptides from Citrullus colocynthis

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1326
Author(s):  
Behzad Shahin-Kaleybar ◽  
Ali Niazi ◽  
Alireza Afsharifar ◽  
Ghorbanali Nematzadeh ◽  
Reza Yousefi ◽  
...  
Keyword(s):  
Trypsin Inhibitor ◽  
High Performance ◽  
De Novo ◽  
Sequence Similarity ◽  
In Silico Analysis ◽  
Reversed Phase ◽  
Amino Acid Sequences ◽  
Peptide Fragments ◽  
Citrullus Colocynthis ◽  
High Sequence Similarity

The plant Citrullus colocynthis, a member of the squash (Cucurbitaceae) family, has a long history in traditional medicine. Based on the ancient knowledge about the healing properties of herbal preparations, plant-derived small molecules, e.g., salicylic acid, or quinine, have been integral to modern drug discovery. Additionally, many plant families, such as Cucurbitaceae, are known as a rich source for cysteine-rich peptides, which are gaining importance as valuable pharmaceuticals. In this study, we characterized the C. colocynthis peptidome using chemical modification of cysteine residues, and mass shift analysis via matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. We identified the presence of at least 23 cysteine-rich peptides in this plant, and eight novel peptides, named citcol-1 to -8, with a molecular weight between ~3650 and 4160 Da, were purified using reversed-phase high performance liquid chromatography (HPLC), and their amino acid sequences were determined by de novo assignment of b- and y-ion series of proteolytic peptide fragments. In silico analysis of citcol peptides revealed a high sequence similarity to trypsin inhibitor peptides from Cucumis sativus, Momordica cochinchinensis, Momordica macrophylla and Momordica sphaeroidea. Using genome/transcriptome mining it was possible to identify precursor sequences of this peptide family in related Cucurbitaceae species that cluster into trypsin inhibitor and antimicrobial peptides. Based on our analysis, the presence or absence of a crucial Arg/Lys residue at the putative P1 position may be used to classify these common cysteine-rich peptides by functional properties. Despite sequence homology and the common classification into the inhibitor cysteine knot family, these peptides appear to have diverse and additional bioactivities yet to be revealed.

scholarly journals Intrachromosomal Recombination within thevsp Locus of Mycoplasma bovis Generates a Chimeric Variable Surface Lipoprotein Antigen

2001 ◽  
Vol 69 (6) ◽  
pp. 3703-3712 ◽  
Author(s):  
Inessa Lysnyansky ◽  
Yael Ron ◽  
Konrad Sachse ◽  
David Yogev
Keyword(s):  
Recombination Event ◽  
Phenotypic Switching ◽  
Upstream Region ◽  
Mycoplasma Bovis ◽  
Gene Repertoire ◽  
Genomic Locus ◽  
Chimeric Genes ◽  
C Terminus ◽  
Variable Surface ◽  
Genes Encoding

ABSTRACT A family of 13 related but divergent vsp genes was recently found in the chromosome of the bovine pathogenMycoplasma bovis. The vsp genomic locus was shown to undergo high-frequency rearrangements and to mediate phenotypic switching of variable lipoprotein antigens (Vsps) on the mycoplasma cell surface. Here we report that the vsp gene repertoire is subject to changes. Genetic analysis of M. bovis clonal isolates displaying distinct Vsp phenotypes showed that an intergenic recombination event between two closely related members of the vsp gene family, the formerly expressedvspA gene and the vspO gene, led to the formation of a new chimeric and functional vsp gene,vspC. The 5′ end of the recombination event was identified within the highly conserved vsp-upstream region, while the 3′ end was localized within the first repetitive domain (RA1) present in both vspA and vspOstructural genes. As a result, the vspC gene is an embodiment of the following domains: an N-terminus-encoding region linked to the highly conserved vsp-upstream region provided by the vspO gene; and a C-terminus-encoding region and the more distal and divergent vsp-upstream region acquired from the vspA gene. The generation of chimeric genes encoding surface antigens may provide an important element of genetic variation and an additional source of antigenic diversification within the mycoplasma population.

scholarly journals Metalloproteinase-like, disintegrin-like, cysteine-rich proteins MDC2 and MDC3: novel human cellular disintegrins highly expressed in the brain

1998 ◽  
Vol 334 (1) ◽  
pp. 93-98 ◽  
Author(s):  
Koji SAGANE ◽  
Yukio OHYA ◽  
Yoshikazu HASEGAWA ◽  
Isao TANAKA
Keyword(s):  
Snake Venom ◽  
Sequence Similarity ◽  
Amino Acid Sequences ◽  
Cytogenet Cell ◽  
Binding Motif ◽  
High Sequence Similarity ◽  
Cell Matrix ◽  
A Disintegrin And Metalloproteinase ◽  
Cell Matrix Interactions ◽  
The Brain

Cellular disintegrins are a family of membrane-anchored proteins structurally related to snake venom disintegrins, and are potential regulators of cell–cell and cell–matrix interactions. The members of this protein family are also called ADAMs (a disintegrin and metalloproteinase) or MDC proteins (metalloproteinase-like disintegrin-like cysteine-rich), because they all contain disintegrin-like and metalloproteinase-like domains. In this paper, we report the cloning and sequence analysis of two novel additional members of this family, which we have termed MDC2 and MDC3. The deduced amino acid sequences reveal that the two proteins possess typical cellular disintegrin structures [that is, pro-, metalloproteinase-like, disintegrin-like, cysteine-rich, epidermal growth factor-like, transmembrane, and cytoplasmic domains] and exhibit high sequence similarity with human MDC/ADAM11 protein [Katagiri, Harada, Emi and Nakamura (1995) Cytogenet. Cell Genet. 68, 39–44]. A zinc-binding motif, which is critical for proteinase activity, is disrupted in the metalloproteinase-like domain of MDC2 and MDC3, as well as MDC/ADAM11. In the disintegrin-like domain of snake venom short disintegrins, the RDG-containing loops are critical for integrin binding. These three MDCs do not contain the RDG sequences, but the corresponding loops in these proteins are similar to each other. Northern blot analysis revealed that the mRNAs of MDC2, MDC3 and MDC/ADAM11 are highly expressed in the brain. These findings suggest that these proteins may function as integrin ligands in the brain.

scholarly journals Extended Repertoire of Genes Encoding Variable Surface Lipoproteins in Mycoplasma bovis Strains

2002 ◽  
Vol 70 (4) ◽  
pp. 2220-2225 ◽  
Author(s):  
Sarit Nussbaum ◽  
Inessa Lysnyansky ◽  
Konrad Sachse ◽  
Sharon Levisohn ◽  
David Yogev
Keyword(s):  
High Frequency ◽  
Antigenic Variation ◽  
Phenotypic Switching ◽  
Mycoplasma Bovis ◽  
Structural Genes ◽  
Coding Sequences ◽  
Sequence Variations ◽  
Variable Surface ◽  
Genes Encoding ◽  
Genomic Cluster

ABSTRACT A genomic cluster of vsp genes was previously shown to mediate high-frequency phenotypic switching of surface lipoprotein antigens in the bovine pathogen Mycoplasma bovis. This study revealed that field strains of M. bovis possess modified versions of the vsp gene complex in which extensive sequence variations occur primarily in the reiterated coding sequences of the vsp structural genes. These findings demonstrate that there is a vastly expanded potential for antigenic variation within populations of this organism.

scholarly journals Antigen heterogeneity among isolates of Mycoplasma bovis is generated by high-frequency variation of diverse membrane surface proteins.

1994 ◽  
Vol 62 (11) ◽  
pp. 5066-5074 ◽  
Author(s):  
R Rosengarten ◽  
A Behrens ◽  
A Stetefeld ◽  
M Heller ◽  
M Ahrens ◽  
...  
Keyword(s):  
High Frequency ◽  
Membrane Surface ◽  
Surface Proteins ◽  
Frequency Variation ◽  
Mycoplasma Bovis

scholarly journals A Putative Multisubunit Na+/H+ Antiporter fromStaphylococcus aureus

1998 ◽  
Vol 180 (24) ◽  
pp. 6642-6648 ◽  
Author(s):  
Toshiaki Hiramatsu ◽  
Kazuyo Kodama ◽  
Teruo Kuroda ◽  
Tohru Mizushima ◽  
Tomofusa Tsuchiya
Keyword(s):  
Sequence Similarity ◽  
Membrane Vesicles ◽  
Amino Acid Sequences ◽  
Open Reading Frames ◽  
Host Cells ◽  
Homology Search ◽  
Upstream Region ◽  
Chromosomal Dna ◽  
E Coli ◽  
Alkaline Conditions

ABSTRACT We cloned several genes encoding an Na+/H+antiporter of Staphylococcus aureus from chromosomal DNA by using an Escherichia coli mutant, lacking all of the major Na+/H+ antiporters, as the host. E. coli cells harboring plasmids for the cloned genes were able to grow in medium containing 0.2 M NaCl (or 10 mM LiCl). Host cells without the plasmids were unable to grow under the same conditions. Na+/H+ antiport activity was detected in membrane vesicles prepared from transformants. We determined the nucleotide sequence of the cloned 7-kbp region. We found that seven open reading frames (ORFs) were necessary for antiporter function. A promoter-like sequence was found in the upstream region from the first ORF. One inverted repeat followed by a T-cluster, which may function as a terminator, was found in the downstream region from the seventh ORF. Neither terminator-like nor promoter-like sequences were found between the ORFs. Thus, it seems that the seven ORFs comprise an operon and that the Na+/H+antiporter consists of seven kinds of subunits, suggesting that this is a novel type of multisubunit Na+/H+antiporter. Hydropathy analysis of the deduced amino acid sequences of the seven ORFs suggested that all of the proteins are hydrophobic. As a result of a homology search, we found that components of the respiratory chain showed sequence similarity with putative subunits of the Na+/H+ antiporter. We observed a large Na+ extrusion activity, driven by respiration in E. coli cells harboring the plasmid carrying the genes. The Na+ extrusion was sensitive to an H+conductor, supporting the idea that the system is not a respiratory Na+ pump but an Na+/H+ antiporter. Introduction of the plasmid into E. coli mutant cells, which were unable to grow under alkaline conditions, enabled the cells to grow under such conditions.

scholarly journals Cloning and characterization of two glutathione S-transferases from a DDT-resistant strain of Anopheles gambiae

1997 ◽  
Vol 324 (1) ◽  
pp. 97-102 ◽  
Author(s):  
Hilary RANSON ◽  
La-aied PRAPANTHADARA ◽  
Janet HEMINGWAY
Keyword(s):  
Anopheles Gambiae ◽  
Resistant Strain ◽  
Sequence Similarity ◽  
Amino Acid Sequences ◽  
Class I ◽  
Kinetic Properties ◽  
Coding Region ◽  
High Sequence Similarity ◽  
Glutathione S Transferases ◽  
High Level

Two cDNA species, aggst1-5 and aggst1-6, comprising the entire coding region of two distinct glutathione S-transferases (GSTs) have been isolated from a 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) resistant strain (ZANDS) of Anopheles gambiae. The nucleotide sequences of these cDNA species share 80.2% identity and their derived amino acid sequences are 82.3% similar. They have been classified as insect class I GSTs on the basis of their high sequence similarity to class I GSTs from Drosophila melanogaster and Musca domestica and they are localized to a region of an An. gambiae chromosome known to contain further class I GSTs. The genes aggst1-5 and aggst1-6 were expressed at high levels in Escherichia coli and the recombinant GSTs were purified by affinity chromatography and characterized. Both agGST1-5 and agGST1-6 showed high activity with the substrates 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene but negligible activity with the mammalian theta class substrates, 1,2-epoxy-3-(4-nitrophenoxy)propane and p-nitrophenyl bromide. Despite their high level of sequence identity, agGST1-5 and agGST1-6 displayed different kinetic properties. Both enzymes were able to metabolize DDT and were localized to a subset of GSTs that, from earlier biochemical studies, are known to be involved in insecticide resistance in An. gambiae. This subset of enzymes is one of three in which the DDT metabolism levels are elevated in resistant insects.

scholarly journals Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment

2007 ◽  
Vol 88 (1) ◽  
pp. 342-350 ◽  
Author(s):  
S. Supyani ◽  
Bradley I. Hillman ◽  
Nobuhiro Suzuki
Keyword(s):  
Active Site ◽  
Sequence Similarity ◽  
Expression System ◽  
Amino Acid Sequences ◽  
Site Directed Mutagenesis ◽  
Sequence Alignments ◽  
High Sequence Similarity ◽  
Chestnut Blight Fungus ◽  
A Genome ◽  
Hypovirulent Strain

The type member Mycoreovirus 1 (MyRV-1) of a newly described genus, Mycoreovirus, isolated from a hypovirulent strain 9B21 of the chestnut blight fungus, has a genome composed of 11 dsRNA segments (S1–S11). All of the segments have single ORFs on their capped, positive-sense strands. Infection of insect cells by baculovirus recombinants carrying full-length cDNAs of S1–S11 resulted in overexpression of protein products of the expected sizes, based on their deduced amino acid sequences. This expression system was utilized to identify the S3-encoded protein (VP3) as a guanylyltransferase by an autoguanylylation assay, in which only VP3 was radiolabelled with [α-32P]GTP. A series of progressive N-terminal and C-terminal deletion mutants was made to localize the autoguanylylation-active site of VP3 to aa residues 133–667. Within this region, a sequence stretch (aa 170–250) with relatively high sequence similarity to homologues of two other mycoreoviruses and two coltiviruses was identified. Site-directed mutagenesis of conserved aa residues revealed that H233, H242, Y243, F244 and F246, but not K172 or K202, play critical roles in guanylyltransferase activities. Together with broader sequence alignments of ‘turreted’ reoviruses, these results supported the a/vxxHx8Hyf/lvf motif, originally noted for orthoreovirus and aquareoviruses, as an active site for guanylyltransferases of viruses within the Orthoreovirus, Aquareovirus, Cypovirus, Oryzavirus, Fijivirus, Coltivirus and Mycoreovirus genera, as well as for the proposed Dinovernavirus genus.

scholarly journals Defensin-like peptide-2 from platypus venom: member of a class of peptides with a distinct structural fold

2000 ◽  
Vol 348 (3) ◽  
pp. 649-656 ◽  
Author(s):  
Allan M. TORRES ◽  
Greg M. DE PLATER ◽  
Magnus DOVERSKOG ◽  
Liesl C. BIRINYI-STRACHAN ◽  
Graham M. NICHOLSON ◽  
...  
Keyword(s):  
Tertiary Structure ◽  
Sequence Similarity ◽  
Three Dimensional ◽  
Amino Acid Sequences ◽  
Dimensional Structure ◽  
High Sequence Similarity ◽  
Natural Function ◽  
Global Fold ◽  
Key Residues ◽  
Β Sheet

The venom of the male Australian duck-billed platypus contains a family of four polypeptides of appox. 5 kDa, which are referred to as defensin-like peptides (DLPs). They are unique in that their amino acid sequences have no significant similarities to those of any known peptides; however, the tertiary structure of one of them, DLP-1, has recently been shown to be similar to β-defensin-12 and to the sodium neurotoxin peptide ShI (Stichodactyla helianthus neurotoxin I). Although DLPs are the major peptides in the platypus venom, little is known about their biological roles. In this study, we determined the three-dimensional structure of DLP-2 by NMR spectroscopy, with the aim of gaining insights into the natural function of the DLPs in platypus venom. The DLP-2 structure was found to incorporate a short helix that spans residues 9-12, and an antiparallel β-sheet defined by residues 15-18 and 37-40. The overall fold and cysteine-pairing pattern of DLP-2 were found to be similar to those of DLP-1, and hence β-defensin-12; however, the sequence similarities between the three molecules are relatively small. The distinct structural fold of the DLP-1, DLP-2, and β-defensin-12 is based upon several key residues that include six cysteines. DLP-3 and DLP-4 are also likely to be folded similarly since they have high sequence similarity with DLP-2. The DLPs, and β-defensin-12 may thus be grouped together into a class of polypeptide molecules which have a common or very similar global fold. The fact that the DLPs did not display antimicrobial, myotoxic, or cell-growth-promoting activities implies that the nature of the side chains in this group of peptides is likely to play an important role in defining the biological function(s).

scholarly journals Multidimensional Analysis of S-alleles from Cross-incompatible Groups of California Almond Cultivars

2006 ◽  
Vol 131 (5) ◽  
pp. 632-636 ◽  
Author(s):  
Kristi K. Barckley ◽  
Sandra L. Uratsu ◽  
Thomas M. Gradziel ◽  
Abhaya M. Dandekar
Keyword(s):  
Sequence Similarity ◽  
Prunus Dulcis ◽  
The United States ◽  
Amino Acid Sequences ◽  
Multidimensional Analysis ◽  
Pairwise Distance ◽  
Self Incompatibility ◽  
High Sequence Similarity ◽  
The World ◽  
S Alleles

The California almond industry is the largest supplier of almonds [Prunus dulcis (Miller) D.A. Webb] in the United States and throughout the world. Self-incompatibility is a major issue in almond production as it greatly affects nut set. In this study, we determined full-length sequences for alleles Sa - Si, determined the genotypes of 44 California cultivars, and assigned the cultivars to cross-incompatibility groups (CIGs). Newly identified S-alleles led to an increase in the number of CIGs. A pairwise distance tree was constructed using the aligned amino acid sequences showing their similarity. Four pairs of alleles (Sc and Se, Sg and Sh, Sd and Sj, and Sb and Sf) showed high sequence similarity. Because of its simplicity, reproducibility, and ease of analysis, PCR is the preferred method for genotyping S-alleles.

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