Amino acid sequences from the N-terminal domain of Bacillus thuringiensis , subspecies alesti , δ-endotoxin

FEBS Letters ◽  
1986 ◽  
Vol 198 (2) ◽  
pp. 283-286 ◽  
Author(s):  
G.G. Chestukhina ◽  
S.A. Tyurin ◽  
A.L. Osterman ◽  
O.P. Khodova ◽  
V.M. Stepanov
Keyword(s):  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Amino Acid Sequences ◽  
Terminal Domain

scholarly journals Sequence Diversity of Bacillus thuringiensis Flagellin (H Antigen) Protein at the Intra-H Serotype Level

2008 ◽  
Vol 74 (17) ◽  
pp. 5524-5532 ◽  
Author(s):  
Dong Xu ◽  
Jean-Charles Côté
Keyword(s):  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Variable Region ◽  
Single Copy ◽  
Amino Acid Sequences ◽  
Sequence Diversity ◽  
Neighbor Joining ◽  
Signature Sequences ◽  
Short Signature ◽  
H Serotypes

ABSTRACT In Bacillus thuringiensis, the hag gene encodes flagellin, the protein responsible for eliciting the immunological reaction in H serotyping. Specific flagellin amino acid sequences have been correlated to specific B. thuringiensis H serotypes, H1 to H67. Ten H serotypes, however, contain three or more antigenic subfactors, labeled a, b, c, d, or e, and have been subdivided into 23 serovars. In the present study, we set out to analyze the sequence diversity of flagellins among serovars from the same H serotypes. We studied the hag genes in 39 B. thuringiensis strains representing the 23 serovars from the 10 H serotypes mentioned above. A serovar and a biovar from an 11th H serotype were also included. The hag genes were amplified and cloned and their nucleotide sequences were determined and translated into amino acid sequences, or the sequences were retrieved directly from GenBank when available. Strains of the H3 serotype contained two or three copies of the fla gene, an ortholog of the hag gene. Strains of the H6 serotype contained three copies. Strains of all other H serotypes each contained a single copy of the hag gene. Alignments of amino acid sequences from all copies in all strains of the H3 serotype revealed short signature sequences, GGAG and SGG, GPDPDDAVKNLT, and DITTTK, that appeared to be specific to the H3c, H3d, and H3e antigenic subfactors, respectively. Similar short signature sequences, GDIT, AFIK, TSAGKA, and SAPSKG, were revealed for H8b, H8c, H20b, and H20c, respectively. Amino acid sequences in the flagellin central variable region were highly conserved among serovars of the H3, H5, H11, and H20 serotypes and much more divergent among serovars of the H4, H10, H18, H24, and H28 serotypes. Two bootstrapped neighbor-joining trees were respectively generated from the alignments of the amino acid sequences translated from all copies of the hag genes in the B. thuringiensis strains of the H3 and H6 serotypes. Sequence identities and relationships were revealed. A third bootstrapped neighbor-joining tree was generated, this one from the alignment of the flagellin amino acid sequences from all the B. thuringiensis strains in the study. Eight clusters, I to VIII, were revealed. Although most clusters contained strains and serovars from the same H serotype, clusters VII and VIII contained serovars from different H serotypes.

scholarly journals Subtypes of NanS-p Sialate O-Acetylesterase Encoded by Stx2a Bacteriophages

Proceedings ◽  
2021 ◽  
Vol 66 (1) ◽  
pp. 15
Author(s):  
Stefanía B. Pascal ◽  
Juan R. Lorenzo Lopez ◽  
Paula M. A. Lucchesi ◽  
Alejandra Krüger
Keyword(s):  
Amino Acid ◽  
Foodborne Pathogens ◽  
Dna Sequences ◽  
Query Sequence ◽  
Amino Acid Sequences ◽  
Bacterial Genomes ◽  
Gene Encoding ◽  
Terminal Domain ◽  
Structural Differences ◽  
Blast Program

Shiga toxin (Stx)-producing Escherichia coli strains are foodborne pathogens that can cause severe human diseases, such as haemorrhagic colitis and haemolytic uraemic syndrome. Stxs are encoded by bacteriophages (Stx phages) which show remarkable variations in genome composition and harbour several genes of unknown function. Recently, a gene encoding a sialate O-acetylesterase (NanS-p) was identified in some relevant Stx2a phages and it was suggested that it could provide advantages for bacterial growth in the gut. The aim of this study was to analyse the presence and sequence of nanS-p genes in available Stx2a phage genomes. A total of 59 DNA sequences of Stx2a phages were extracted from the NCBI GenBank database with the BLAST program using the stx2a sequence from phage 933W as a query sequence, either as complete phage genomes (45) or from bacterial genomes by subsequent analysis with the PHASTER web server (14). Comparative analysis revealed that nanS-p was located downstream of stx2a in all genomes. Twenty different amino acid sequences of NanS-p were identified. Specifically, catalytic esterase domains showed only 11 possible sequences, with differences mainly observed in nine amino acid positions. Sequences corresponding to the N-terminal domain (DUF1737) showed three possible sequences, two of them closely related, while the C-terminal domain was highly variable, with four groups with structural differences. Since sialate O-acetylesterase activity has been determined from particular Stx2a phages, new studies are necessary to evaluate if the NanS-p subtypes identified in the present study also differ in their biological activity.

Y-box proteins combine versatile cold shock domains and arginine-rich motifs (ARMs) for pleiotropic functions in RNA biology

2018 ◽  
Vol 475 (17) ◽  
pp. 2769-2784 ◽  
Author(s):  
Kenneth C. Kleene
Keyword(s):  
Amino Acid ◽  
Cold Shock ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Specific Binding ◽  
Amino Acid Sequences ◽  
Site Specific ◽  
Terminal Domain ◽  
Rna Backbone ◽  
Specific Regulation

Y-box proteins are single-strand DNA- and RNA-binding proteins distinguished by a conserved cold shock domain (CSD) and a variable C-terminal domain organized into alternating short modules rich in basic or acidic amino acids. A huge literature depicts Y-box proteins as highly abundant, staggeringly versatile proteins that interact with all mRNAs and function in most forms of mRNA-specific regulation. The mechanisms by which Y-box proteins recognize mRNAs are unclear, because their CSDs bind a jumble of diverse elements, and the basic modules in the C-terminal domain are considered to bind nonspecifically to phosphates in the RNA backbone. A survey of vertebrate Y-box proteins clarifies the confusing names for Y-box proteins, their domains, and RNA-binding motifs, and identifies several novel conserved sequences: first, the CSD is flanked by linkers that extend its binding surface or regulate co-operative binding of the CSD and N-terminal and C-terminal domains to proteins and RNA. Second, the basic modules in the C-terminal domain are bona fide arginine-rich motifs (ARMs), because arginine is the predominant amino acid and comprises 99% of basic residues. Third, conserved differences in AA (amino acid) sequences between isoforms probably affect RNA-binding specificity. C-terminal ARMs connect with many studies, demonstrating that ARMs avidly bind sites containing specific RNA structures. ARMs crystallize insights into the under-appreciated contributions of the C-terminal domain to site-specific binding by Y-box proteins and difficulties in identifying site-specific binding by the C-terminal domain. Validated structural biology techniques are available to elucidate the mechanisms by which YBXprot (Y-box element-binding protein) CSDs and ARMs identify targets.

scholarly journals Cry1Aa binding to the cadherin receptor does not require conserved amino acid sequences in the domain II loops

2012 ◽  
Vol 33 (1) ◽  
Author(s):  
Yuki Fujii ◽  
Shiho Tanaka ◽  
Manami Otsuki ◽  
Yasushi Hoshino ◽  
Chinatsu Morimoto ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
No Reduction ◽  
Amino Acid Sequences ◽  
Binding Affinities ◽  
Binding Mechanism ◽  
Wild Type ◽  
Critical Amino Acid ◽  
Cry Toxin

Characterizing the binding mechanism of Bt (Bacillus thuringiensis) Cry toxin to the cadherin receptor is indispensable to understanding the specific insecticidal activity of this toxin. To this end, we constructed 30 loop mutants by randomly inserting four serial amino acids covering all four receptor binding loops (loops α8, 1, 2 and 3) and analysed their binding affinities for Bombyx mori cadherin receptors via Biacore. High binding affinities were confirmed for all 30 mutants containing loop sequences that differed from those of wild-type. Insecticidal activities were confirmed in at least one mutant from loops 1, 2 and 3, suggesting that there is no critical amino acid sequence for the binding of the four loops to BtR175. When two mutations at different loops were integrated into one molecule, no reduction in binding affinity was observed compared with wild-type sequences. Based on these results, we discussed the binding mechanism of Cry toxin to cadherin protein.

scholarly journals Molecular cloning, expression and regulatory activity of Gα11- and βγ-subunit-stimulated phospholipase C-β from avian erythrocytes

1996 ◽  
Vol 316 (2) ◽  
pp. 559-568 ◽  
Author(s):  
Gary L. WALDO ◽  
Andrew PATERSON ◽  
José L. BOYER ◽  
Robert A. NICHOLAS ◽  
T. Kendall HARDEN
Keyword(s):  
Amino Acid ◽  
Phospholipase C ◽  
G Protein ◽  
Recombinant Baculovirus ◽  
Recombinant Enzyme ◽  
Amino Acid Sequences ◽  
Calculated Molecular Mass ◽  
Reading Frame ◽  
Terminal Domain ◽  
Α Subunits

A turkey erythrocyte phospholipase C (PLC) has been instrumental in delineating the role of G-proteins in receptor-regulated inositol lipid signalling. This isoenzyme is uniquely regulated both by α-subunits of the Gq family and by G-protein βγ-subunits. A 4819 bp cDNA encoding this PLC has been cloned from a turkey erythrocyte cDNA library. The open reading frame of this cDNA encodes a 1211-amino-acid protein (calculated molecular mass 139050 Da) that contains amino acid sequences of 16 peptides sequenced from the turkey erythrocyte PLC. The predicted sequence of the turkey PLC shows considerable similarity with the sequences of previously cloned members of the PLC-β family, with the highest identity (71%) shared with PLC-β2 and lesser identities observed with PLC-β1 (49%), PLC-β3 (46%) and PLC-β4 (37%). The largest differences in sequence between the turkey PLC-β and other PLC-β isoenzymes occur in the C-terminal domain and in the region between the X- and Y-domains. The turkey isoenzyme and PLC-β2, which differ in their regulation by G-protein α-subunits, are only 44% similar across the approx. 400 amino acid residues of the C-terminal domain that has been implicated in αq activation of these proteins. Recombinant turkey PLC-β was purified to homogeneity following expression from a recombinant baculovirus in Sf9 insect cells. The immunoreactivity and mobility on SDS/PAGE of the recombinant enzyme were the same as observed with native turkey erythrocyte PLC-β. Moreover, the catalytic activities of the recombinant enzyme were indistinguishable from those of native turkey erythrocyte PLC-β in assays carried out in the presence of cholate and Ca2+, or in assays of activity after reconstitution with Gα11 or G-protein βγ-subunits. The turkey PLC-β was more sensitive to activation by Gα11 than was PLC-β2, and was more sensitive to activation by βγ-subunits than either PLC-β2 or PLC-β1.

scholarly journals Distinct clpP Genes Control Specific Adaptive Responses in Bacillus thuringiensis

2002 ◽  
Vol 184 (20) ◽  
pp. 5554-5562 ◽  
Author(s):  
Sinda Fedhila ◽  
Tarek Msadek ◽  
Patricia Nel ◽  
Didier Lereclus
Keyword(s):  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Low Temperature ◽  
Bacterial Species ◽  
Growth Cycle ◽  
Amino Acid Sequences ◽  
Adaptive Responses ◽  
Regulatory Pathways ◽  
Eukaryotic Organisms ◽  
Mutant Cells

ABSTRACT ClpP and ClpC are subunits of the Clp ATP-dependent protease, which is ubiquitous among prokaryotic and eukaryotic organisms. The role of these proteins in stress tolerance, stationary-phase adaptive responses, and virulence in many bacterial species has been demonstrated. Based on the amino acid sequences of the Bacillus subtilis clpC and clpP genes, we identified one clpC gene and two clpP genes (designated clpP1 and clpP2) in Bacillus thuringiensis. Predicted proteins ClpP1 and ClpP2 have approximately 88 and 67% amino acid sequence identity with ClpP of B. subtilis, respectively. Inactivation of clpC in B. thuringiensis impaired sporulation efficiency. The clpP1 and clpP2 mutants were both slightly susceptible to salt stress, whereas disruption of clpP2 negatively affected sporulation and abolished motility. Virulence of the clp mutants was assessed by injecting bacteria into the hemocoel of Bombyx mori larvae. The clpP1 mutant displayed attenuated virulence, which appeared to be related to its inability to grow at low temperature (25°C), suggesting an essential role for ClpP1 in tolerance of low temperature. Microscopic examination of clpP1 mutant cells grown at 25°C showed altered bacterial division, with cells remaining attached after septum formation. Analysis of lacZ transcriptional fusions showed that clpP1 was expressed at 25 and 37°C during the entire growth cycle. In contrast, clpP2 was expressed at 37°C but not at 25°C, suggesting that ClpP2 cannot compensate for the absence of ClpP1 in the clpP1 mutant cells at low temperature. Our study demonstrates that ClpP1 and ClpP2 control distinct cellular regulatory pathways in B. thuringiensis.

scholarly journals Sequence Diversity of the Bacillus thuringiensis and B. cereus Sensu Lato Flagellin (H Antigen) Protein: Comparison with H Serotype Diversity

2006 ◽  
Vol 72 (7) ◽  
pp. 4653-4662 ◽  
Author(s):  
Dong Xu ◽  
Jean-Charles Côté
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Nucleotide Sequences ◽  
Amino Acid Sequences ◽  
Sequence Diversity ◽  
The Other ◽  
Bacillus Halodurans ◽  
V Region ◽  
H Serotypes

ABSTRACT We set out to analyze the sequence diversity of the Bacillus thuringiensis flagellin (H antigen [Hag]) protein and compare it with H serotype diversity. Some other Bacillus cereus sensu lato species and strains were added for comparison. The internal sequences of the flagellin (hag) alleles from 80 Bacillus thuringiensis strains and 16 strains from the B. cereus sensu lato group were amplified and cloned, and their nucleotide sequences were determined and translated into amino acids. The flagellin allele nucleotide sequences for 10 additional strains were retrieved from GenBank for a total of 106 Bacillus species and strains used in this study. These included 82 B. thuringiensis strains from 67 H serotypes, 5 B. cereus strains, 3 Bacillus anthracis strains, 3 Bacillus mycoides strains, 11 Bacillus weihenstephanensis strains, 1 Bacillus halodurans strain, and 1 Bacillus subtilis strain. The first 111 and the last 66 amino acids were conserved. They were referred to as the C1 and C2 regions, respectively. The central region, however, was highly variable and is referred to as the V region. Two bootstrapped neighbor-joining trees were generated: a first one from the alignment of the translated amino acid sequences of the amplified internal sequences of the hag alleles and a second one from the alignment of the V region amino acid sequences, respectively. Of the eight clusters revealed in the tree inferred from the entire C1-V-C2 region amino acid sequences, seven were present in corresponding clusters in the tree inferred from the V region amino acid sequences. With regard to B. thuringiensis, in most cases, different serovars had different flagellin amino acid sequences, as might have been expected. Surprisingly, however, some different B. thuringiensis serovars shared identical flagellin amino acid sequences. Likewise, serovars from the same H serotypes were most often found clustered together, with exceptions. Indeed, some serovars from the same H serotype carried flagellins with sufficiently different amino acid sequences as to be located on distant clusters. Species-wise, B. halodurans, B. subtilis, and B. anthracis formed specific branches, whereas the other four species, all in the B. cereus sensu lato group, B. mycoides, B. weihenstephanensis, B. cereus, and B. thuringiensis, did not form four specific clusters as might have been expected. Rather, strains from any of these four species were placed side by side with strains from the other species. In the B. cereus sensu lato group, B. anthracis excepted, the distribution of strains was not species specific.

Characterization ofBacillus thuringiensissubsp.kurstakistrain S93 effective against the fall armywormSpodoptera frugiperda)

1999 ◽  
Vol 45 (6) ◽  
pp. 464-471 ◽  
Author(s):  
Joseilde O Silva-Werneck ◽  
Marlene T De-Souza ◽  
José MC de S. Dias ◽  
Bergmann M Ribeiro
Keyword(s):  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Spodoptera Frugiperda ◽  
Fall Armyworm ◽  
Amino Acid Sequences ◽  
Dna Library ◽  
Sds Page ◽  
Total Dna ◽  
Type Gene ◽  
High Degree

A Brazilian strain of Bacillus thuringiensis subsp. kurstaki, designated S93, was analyzed regarding its cry gene and protein contents and activity against the fall armyworm (Spodoptera frugiperda, Smith 1797). Bioassays using lyophilized powders of S93 or HD-1 and third instar larvae of S. frugiperda showed a 12.3-fold lower LC50for the S93 strain when compared with the standard HD-1 strain. The spore-crystal mixture, analyzed by SDS-PAGE, showed two major polypeptides of 130 and 65 kDa, corresponding to Cry1 and Cry2 toxins, respectively. Western blot analysis showed that these proteins were immunologically related to the Cry1A protein from B. thuringiensis subsp. kurstaki HD-73. The polymerase chain reaction technique (PCR) using total DNA from the S93 strain and specific primers showed the presence of cry1Aa, cry1Ab, and cry1Ac genes, and a cry1A-type gene was localized in a plasmid of about 44 MDa. A cry1Ab gene was isolated from a S93 plasmid DNA library and completely sequenced. Computer analysis showed that the gene sequence (GenBank acession number AF059670) is identical to cry1Ab1 and has 91.6 and 85.9% identity with cry1Aa1 and cry1Ac1 genes, respectively. The deduced amino-acid sequence showed a high degree of similarity with the amino-acid sequences of the Cry1Ab1 (100%), Cry1Aa1 (93.8%), and Cry1Ac1 (90.6%) proteins.Key words: Bacillus thuringiensis, Spodoptera frugiperda, biological control, crystal protein, cry genes.

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