158. Adenovirus-Mediated Delivery of a Y. pestis V Antigen Variant with Reduced Immunomodulatory Properties Elicits Comparable Serum Anti-V Antigen IgG Titers to Adenovirus-Mediated Delivery of Wild-Type V Antigen

The lethal effects of the expression of the oncogenic protein tyrosine kinase p60v-src in Saccharomyces cerevisiae are associated with a loss of cell cycle control at the G1/S and G2/M checkpoints. Results described here indicate that the ability of v-Src to kill yeast is dependent on the integrity of the SH2 domain, a region of the Src protein involved in recognition of proteins phosphorylated on tyrosine. Catalytically active v-Src proteins with deletions in the SH2 domain have little effect on yeast growth, unlike wild-type v-Src protein, which causes accumulation of large-budded cells, perturbation of spindle microtubules and increased DNA content when expressed. The proteins phosphorylated on tyrosine in cells expressing v-Src differ from those in cells expressing a Src protein with a deletion in the SH2 domain. Also, unlike the wild-type v-Src protein, which drastically increases histone H1-associated Cdc28 kinase activity, c-Src and an altered v-Src protein have no effect on Cdc28 kinase activity. These results indicate that the SH2 domain is functionally important in the disruption of the yeast cell cycle by v-Src.

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Lipid profiles of conidia of Aspergillus niger and a fatty acid auxotroph

Canadian Journal of Microbiology ◽

10.1139/m87-195 ◽

1987 ◽

Vol 33 (12) ◽

pp. 1116-1120 ◽

Cited By ~ 8

Author(s):

Panchanon Chattopadhyay ◽

Santu Kumar Banerjee ◽

Kalyani Sen ◽

Parul Chakrabarti

Keyword(s):

Fatty Acids ◽

Fatty Acid ◽

Double Bond ◽

Aspergillus Niger ◽

Fatty Acid Profile ◽

Unsaturated Fatty Acid ◽

Desaturase Activity ◽

Wild Type ◽

Type V ◽

Fatty Acid Auxotroph

Conidial lipids of the wild-type (V35) Aspergillus niger and its unsaturated fatty acid auxotroph (UFA2) were compared. The wild type contained lower levels (7.6%) of phospholipids and higher levels (28.4%) of glycolipids than the mutant (16.5 and 22.2%, respectively). Oleic (33.4%), linoleic (22.5%), palmitic (12.8%), stearic (7.4%), and linolenic (6.2%) were the main fatty acids of the wild type (V35). The mutant grew only in the presence of unsaturated fatty acid having at least one Δ9cis double bond, and its conidial fatty acid profile was influenced by the exogenous acid. Analyses of the fatty acids of UFA2 grown in the presence of different fatty acid supplements support the original view that the mutant is defective in Δ9-desaturase activity.

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Identification of nonessential disulfide bonds and altered conformations in the v-sis protein, a homolog of the B chain of platelet-derived growth factor.

Molecular and Cellular Biology ◽

10.1128/mcb.8.3.1011 ◽

1988 ◽

Vol 8 (3) ◽

pp. 1011-1018 ◽

Cited By ~ 13

Author(s):

M K Sauer ◽

D J Donoghue

Keyword(s):

Biological Activity ◽

Growth Factor ◽

Disulfide Bonds ◽

Polyacrylamide Gel Electrophoresis ◽

Protein A ◽

Sodium Dodecyl ◽

Biologically Active ◽

Platelet Derived Growth Factor ◽

Wild Type ◽

Type V

The protein encoded by v-sis, the oncogene of simian sarcoma virus, is homologous to the B chain of platelet-derived growth factor (PDGF). There are eight conserved Cys residues between PDGF-B and the v-sis protein. Both native PDGF and the v-sis protein occur as disulfide-bonded dimers, probably containing both intramolecular and intermolecular disulfide bonds. Oligonucleotide-directed mutagenesis was used to change the Cys codons to Ser codons in the v-sis gene. Four single mutants lacked detectable biological activity, indicating that Cys-127, Cys-160, Cys-171, and Cys-208 are required for formation of a biologically active v-sis protein. The other four single mutants retained biological activity as determined in transformation assays, indicating that Cys-154, Cys-163, Cys-164, and Cys-210 are dispensable for biological activity. Double and triple mutants containing three of these altered sites were constructed, some of which were transforming as well. The v-sis proteins encoded by biologically active mutants displayed significantly reduced levels of dimeric protein compared with the wild-type v-sis protein, which dimerized very efficiently. Furthermore, a mutant with a termination codon at residue 209 exhibited partial transforming activity. This study thus suggests that the minimal region required for transformation consists of residues 127 to 208. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the v-sis proteins encoded by some of the biologically active mutants exhibited an altered conformation when compared with the wild-type v-sis protein, and suggested that Cys-154 and Cys-163 participate in a nonessential disulfide bond.

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Characterization of BcaA, a Putative Classical Autotransporter Protein in Burkholderia pseudomallei

Infection and Immunity ◽

10.1128/iai.01453-12 ◽

2013 ◽

Vol 81 (4) ◽

pp. 1121-1128 ◽

Cited By ~ 9

Author(s):

Cristine G. Campos ◽

Luke Borst ◽

Peggy A. Cotter

Keyword(s):

Burkholderia Pseudomallei ◽

Efflux Pump ◽

Outer Membrane Proteins ◽

Lung Epithelial Cells ◽

Wild Type ◽

Passenger Domain ◽

Content Type ◽

Reading Frame ◽

Type V

ABSTRACTBurkholderia pseudomalleiis a tier 1 select agent, and the causative agent of melioidosis, a disease with effects ranging from chronic abscesses to fulminant pneumonia and septic shock, which can be rapidly fatal. Autotransporters (ATs) are outer membrane proteins belonging to the type V secretion system family, and many have been shown to play crucial roles in pathogenesis. The open reading frame Bp1026b_II1054 (bcaA) inB. pseudomalleistrain 1026b is predicted to encode a classical autotransporter protein with an approximately 80-kDa passenger domain that contains a subtilisin-related domain. Immediately 3′ tobcaAis Bp11026_II1055 (bcaB), which encodes a putative prolyl 4-hydroxylase. To investigate the role of these genes in pathogenesis, large in-frame deletion mutations ofbcaAandbcaBwere constructed in strain Bp340, an efflux pump mutant derivative of the melioidosis clinical isolate 1026b. Comparison of Bp340ΔbcaAand Bp340ΔbcaBmutants to wild-typeB. pseudomalleiin vitrodemonstrated similar levels of adherence to A549 lung epithelial cells, but the mutant strains were defective in their ability to invade these cells and to form plaques. In a BALB/c mouse model of intranasal infection, similar bacterial burdens were observed after 48 h in the lungs and liver of mice infected with Bp340ΔbcaA, Bp340ΔbcaB, and wild-type bacteria. However, significantly fewer bacteria were recovered from the spleen of Bp340ΔbcaA-infected mice, supporting the idea of a role for this AT in dissemination or in survival in the passage from the site of infection to the spleen.

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Requirement for an Interaction of XRCC4 with DNA Ligase IV for Wild-type V(D)J Recombination and DNA Double-strand Break Repairin Vivo

Journal of Biological Chemistry ◽

10.1074/jbc.273.38.24708 ◽

1998 ◽

Vol 273 (38) ◽

pp. 24708-24714 ◽

Cited By ~ 108

Author(s):

Ulf Grawunder ◽

David Zimmer ◽

Peter Kulesza ◽

Michael R. Lieber

Keyword(s):

Strand Break ◽

Double Strand Break ◽

Dna Ligase ◽

Wild Type ◽

Dna Ligase Iv ◽

Dna Double Strand Break ◽

Ligase Iv ◽

Type V

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Identification of nonessential disulfide bonds and altered conformations in the v-sis protein, a homolog of the B chain of platelet-derived growth factor

Molecular and Cellular Biology ◽

10.1128/mcb.8.3.1011-1018.1988 ◽

1988 ◽

Vol 8 (3) ◽

pp. 1011-1018

Author(s):

M K Sauer ◽

D J Donoghue

Keyword(s):

Biological Activity ◽

Growth Factor ◽

Disulfide Bonds ◽

Polyacrylamide Gel Electrophoresis ◽

Protein A ◽

Sodium Dodecyl ◽

Biologically Active ◽

Platelet Derived Growth Factor ◽

Wild Type ◽

Type V

The protein encoded by v-sis, the oncogene of simian sarcoma virus, is homologous to the B chain of platelet-derived growth factor (PDGF). There are eight conserved Cys residues between PDGF-B and the v-sis protein. Both native PDGF and the v-sis protein occur as disulfide-bonded dimers, probably containing both intramolecular and intermolecular disulfide bonds. Oligonucleotide-directed mutagenesis was used to change the Cys codons to Ser codons in the v-sis gene. Four single mutants lacked detectable biological activity, indicating that Cys-127, Cys-160, Cys-171, and Cys-208 are required for formation of a biologically active v-sis protein. The other four single mutants retained biological activity as determined in transformation assays, indicating that Cys-154, Cys-163, Cys-164, and Cys-210 are dispensable for biological activity. Double and triple mutants containing three of these altered sites were constructed, some of which were transforming as well. The v-sis proteins encoded by biologically active mutants displayed significantly reduced levels of dimeric protein compared with the wild-type v-sis protein, which dimerized very efficiently. Furthermore, a mutant with a termination codon at residue 209 exhibited partial transforming activity. This study thus suggests that the minimal region required for transformation consists of residues 127 to 208. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the v-sis proteins encoded by some of the biologically active mutants exhibited an altered conformation when compared with the wild-type v-sis protein, and suggested that Cys-154 and Cys-163 participate in a nonessential disulfide bond.

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Wild-type V(D)J recombination in scid pre-B cells

Molecular and Cellular Biology ◽

10.1128/mcb.10.10.5397-5407.1990 ◽

1990 ◽

Vol 10 (10) ◽

pp. 5397-5407

Author(s):

E A Hendrickson ◽

M S Schlissel ◽

D T Weaver

Keyword(s):

B Cells ◽

Cell Clone ◽

Receptor Gene ◽

Cell Receptor ◽

Lymphoid Cells ◽

Homozygous Mutation ◽

Wild Type ◽

Functional Protein ◽

Cell Clones ◽

Type V

Homozygous mutation at the scid locus in the mouse results in the aberrant rearrangement of immunoglobulin and T-cell receptor gene segments. We introduced a retroviral vector containing an inversional immunoglobulin rearrangement cassette into scid pre-B cells. Most rearrangements were accompanied by large deletions, consistent with previously characterized effects of the scid mutation. However, two cell clones were identified which contained perfect reciprocal fragments and wild-type coding joints, documenting, on a molecular level, the ability of scid pre-B cells to generate functional protein-coding domains. Subsequent rearrangement of the DGR cassette in one of these clones was accompanied by a deletion, suggesting that this cell clone had not reverted the scid mutation. Indeed, induced rearrangement of the endogenous kappa loci in these two cell clones resulted in a mixture of scid and wild-type V-J kappa joints, as assayed by a polymerase chain reaction and DNA sequencing. In addition, three immunoglobulin mu- scid pre-B cell lines showed both scid and wild-type V-J kappa joins. These experiments strongly suggest that the V(D)J recombinase activity in scid lymphoid cells is diminished but not absent, consistent with the known leakiness of the scid mutation.

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A modifier gene involved in the expression of the dominant mating type allele in Paramecium caudatum

Genetics Research ◽

10.1017/s0016672300016347 ◽

1976 ◽

Vol 27 (2) ◽

pp. 135-141 ◽

Cited By ~ 4

Author(s):

Koichi Hiwatashi ◽

Koji Myohara

Keyword(s):

Mating Type ◽

Modifier Gene ◽

Recessive Allele ◽

Dominant Allele ◽

Paramecium Caudatum ◽

Mutant Clone ◽

Wild Type ◽

Type Allele ◽

Type V

SUMMARYMating type in Paramecium caudatum, syngen 3 is determined by a pair of alleles with simple dominance; the recessive allele restricts homozygotes to mating type V and the dominant allele permits expression of mating type VI. Clones of mating type V never show natural selfing, but most clones of mating type VI self naturally. A mutant clone of mating type VI which never selfed over a period of more than 3 years was obtained by treatment with N-methyl-N′-nitro-N-nitrosoguanidine. When this mutant clone was crossed to a wild-type stock of mating type V, all F1 clones of mating type VI gave rise to selfers. From selfing of these F1 of mating type VI, clones of F2 were obtained. Nearly 3:1 segregation of selfer to non-selfer clones was observed among the F2 clones of mating type VI. The results were consistent with the interpretation that a dominant modifier gene, Su(+mtV), controls the instability in the expression of mating type VI.

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