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PubMed2006

Effect of the morphogene bolA on the permeability of the Escherichia coli outer membrane.

Escherichia coli bolA is a morphogene involved in stress response and cell division. Overexpression of bolA induces biofilm formation and affects the levels of carboxypeptidases PBP5, PBP6 and beta-lactamase AmpC. In this study, we have shown that changes in the expression of bolA result in alterations in the properties of the outer membrane. The sensitivity to detergents and vancomycin was reduced when bolA was overexpressed and fluorescent probes indicated that different levels of bolA had an effect on outer membrane protein accessibility. Moreover, bolA was shown to be involved in the modulation of the OmpF/OmpC balance.

Bacterial Outer Membrane Proteins↗
PubMed1989

The influence of the BoLA-A locus on reproductive traits in cattle.

Associations between the major histocompatibility complex (MHC) and reproductive performance have been reported in humans, mice, rats, pigs and chickens. Only the A locus of the bovine major histocompatibility complex (BoLA-A) has been well characterized, and 42 alleles of this locus have been identified in American cattle. Four studies were conducted to examine the association between alleles of the BoLA-A locus and reproductive performance. Testis size, which is an indicator of early puberty and increased fertility in young bulls, was examined in 440 yearling bulls from nine breeds with a gene substitution model that included the effects of breed, sire, age of dam and age or weight of the bull. Estimated breeding value for twinning was examined with a gene substitution model with 204 cattle from a herd with a high frequency of twinning. Fertility of potential partners having BoLA-A locus alleles in common was examined in a prospective study involving 101 pure-bred Hereford cows mated by artificial insemination to four pure-bred Hereford bulls. The effect of homozygosity on birth weight, preweaning weight gain and post-weaning weight gain was estimated in a sample of 683 calves from nine breeds; 22% of the calves were apparently homozygous and 78% were heterozygous at the BoLA-A locus. There were significant and large effects of some BoLA-A locus alleles on paired testicular volume, but the analyses on the other traits did not show significant associations. Substitution of the W6.1 allele for the W9A allele reduced paired testicular volume by 150 +/- 44 cm3. The W6.1 allele has now been shown to influence a reproductive trait, a production trait and susceptibility to an economically important disease. Selection for these traits may influence the frequency of the large number of alleles at the BoLA-A locus.

Alleles↗
PubMed1989

Induction of a growth-phase-dependent promoter triggers transcription of bolA, an Escherichia coli morphogene.

The bolA gene, which is involved in the morphogenetic pathways of Escherichia coli, was sequenced and two potential promoters were identified. Expression from promoter P1, proximal to the bolA structural gene is specifically induced during the transition to the stationary phase of growth. This promoter contains an unusual--10 region (CGGCTAGTA), which defines a new class of E. coli promoters necessary for the dramatic increase in the rate of synthesis of a large set of proteins during the cessation of logarithmic growth. This conclusion was confirmed by identifying two additional E. coli promoters and one plasmid promoter, which also were induced during the transition to the stationary phase of growth. Analysis of proteins produced during the exponential and stationary phases of growth in a bolA null mutant suggest a possible role for the BolA protein in the induction of the expression of penicillin-binding protein 6 (PBP6) in the transition to the stationary phase. Supporting this hypothesis is the presence of a putative DNA-binding domain within the bolA coding sequence.

Amino Acid Sequence↗
PubMed1981

The bovine major histocompatibility complex (BoLa): close linkage of the genes controlling serologically defined antigens and mixed lymphocyte reactivity.

Detection of linkage between genetic loci in cattle has been hampered by the lack of large full -sib families. A unique source of full-sib families is now available from embryo transplantation. Lymphocytes from six full-sib families, ranging in size from three to seven siblings, were tested for serologically defined BoLA antigens (BoLA-A). In addition, mixed lymphocyte reactivity (MLR) was tested between all paired combinations of cells within each family to distinguish BoLA-D specificities. Serologically identical siblings within each family were reciprocally nonreactive in MLR, and vice versa; thus, no recombinants were detected between the BoLA-A and the BoLA-D loci. Classical genetic linkage analysis revealed that these loci are significantly closer than 11.9 centimorgans.

Animals↗
PubMed1999

Evidence for an additional cattle DQA locus, BoLA-DQA5.

Studying the genetic polymorphism of the major histocompatibility complex class II genes in cattle, we identified an allele (BNI13) which encodes a typical class II alpha chain. Its transcription was confirmed by RNA analysis. Sequence comparisons, Southern blot, and phylogenetic analyses indicate that (1) BNI13 represents a distinct DQA locus which we propose to designate BoLA-DQA5, (2) BoLA-DQA1 and BoLA-DQA5 separated after the divergence of BoLA-DQA1 and BoLA-DQA2, but prior to the separation of sheep DQA1 and cattle DQA1, and (3) DQA5 is distributed among various cattle breeds but is confined to certain haplotypes.

Alleles↗
PubMed1992

Mapping of bovine markers CYP21, PRL, and BOLA DRBP1 by genetic linkage analysis in reference pedigrees.

We have analyzed DNA from 13 bovine reference pedigrees using primers specific for microsatellite markers derived from the 21-steroid hydroxylase (CYP21) and prolactin (PRL) genes and the leukocyte antigen (BOLA DRBP1) pseudogene. Linkage was demonstrated between PRL and BOLA DRBP1 (theta = 0.05; Z = 19.6), cyp21 and PRL (theta = 0.13; Z = 6.8), and BOLA DRBP1 and CYP21 (theta = 0.17; Z = 10.4). These results suggest an order BOLA DRBP1-PRL-CYP21, although in a multilocus analysis the alternative order PRL-BOLA DRBP1-CYP21 was also possible. The data confirm and extend the previously established syntenic relationship between these markers on bovine chromosome 23 and provide points of anchorage for further linkage studies in the reference pedigrees described.

Animals↗
PubMed2004

Gene frequency distribution of the BoLA-DRB3 locus in Saavedreño Creole dairy cattle.

The objective of this study is to describe the gene frequency distribution of the bovine lymphocyte antigen (BoLA)-DRB3 locus in Saavedreño Creole dairy cattle and to compare it with previously reported patterns in other cattle breeds. One hundred and twenty-five Saavedreño Creole dairy cattle were genotyped for the BoLA-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism. Twenty-two out of 53 previously identified BoLA-DRB3.2 alleles were detected, with gene frequencies ranging from 0.4 to 16.8%. Seventy percent of the variation corresponded to the seven most frequent alleles (BoLA-DRB3.2*7, *8, *11, *16, *27, *36, and *37). The studied population exhibits a high degree of expected heterozygosity (he = 0.919). The FIS index did not show significant deviation from Hardy-Weinberg equilibrium. However, the neutrality test showed an even gene frequency distribution. This result could be better explained assuming balancing selection instead of neutral or positive selection for one or a few alleles. In conclusion, the results of this study demonstrated that BoLA-DRB3.2 is a highly polymorphic locus in Saavedreño Creole dairy cattle, with significant variation in allele frequency among cattle breeds.

Alleles↗
PubMed2002

The gene bolA regulates dacA (PBP5), dacC (PBP6) and ampC (AmpC), promoting normal morphology in Escherichia coli.

The gene bolA has been shown to trigger the formation of osmotically stable round cells when overexpressed in stationary phase. We show that in poor growth conditions bolA is essential for normal cell morphology in stationary phase and under conditions of starvation. During exponential growth bolA promotes round morphology through a mechanism that is strictly dependent on the two main Escherichia colid,d-carboxypeptidases, PBP5 and PBP6. The results show that bolA controls the levels of transcription of dacA (PBP5), dacC (PBP6) and ampC (AmpC), a class C beta-lactamase, thus connecting for the first time penicillin binding proteins (PBPs) and beta-lactamases at the level of gene regulation. Furthermore, PBP5 and PBP6 are shown to be independently regulated and to have distinct effects on the peptidoglycan layer. The evidence presented demonstrates that bolA is a regulator of cell wall biosynthetic enzymes with different roles in cell morphology and cell division.

Bacterial Proteins↗
PubMed1987

Expression and genetic segregation of parental BoLA serotypes in bovine embryos.

The detection of parentally derived BoLA serotypes was attempted in 68 bovine embryos. 23 bovine embryos were tested for genetic segregation of maternally derived BoLA serotypes. 45 bovine embryos were tested for genetic segregation of paternally derived BoLA serotypes. The expected parentally derived BoLA gene products were detectable on approximately 50% of the embryos tested. A 1:1 segregation ratio of expression or non-expression of parental BoLA serotypes in 7-day-old preimplantation bovine embryos, which is expected for codominant alleles, could not be rejected.

Animals↗
PubMed1989

Joint report of the Third International Bovine Lymphocyte Antigen (BoLA) Workshop, Helsinki, Finland, 27 July 1986.

Two hundred and eighty-two alloantisera were submitted by 20 participating laboratories from 13 countries and tested against lymphocytes of 1298 cattle. The cell panel consisted of samples from 38 Bos taurus breeds, 11 Bos taurus crossbreeds, 4 Bos indicus breeds, 6 Bos taurus x Bos indicus, and a variety of other crossbred populations. Using a standardized lymphocytotoxicity test, all 17 previously identified BoLA specificities were confirmed. The workshop produced agreement on 16 new lymphocyte alloantigenic specificities. Three of the new specificities behaved as splits of previously identified BoLA specificities. Four of the new specificities behaved as alleles at the agreed BoLA-A locus. Seven new specificities are tentatively assigned to the BoLA-A locus but require further definition. Two new specificities may represent products of a second closely-linked BoLA locus.

Animals↗
PubMed1992

Extensive polymorphism of the BoLA-DRB3 gene distinguished by PCR-RFLP.

A polymerase chain reaction (PCR)-based method is described for typing of alleles of the bovine lymphocyte antigen (BoLA)-DRB3 gene. A total of 30 DRB3 alleles were distinguished by digestion of PCR amplification products of BoLA-DRB3 exon 2 with RsaI, BstYI and HaeIII (PCR-RFLP). All restriction fragment patterns, with the exception of one HaeIII pattern, were consistent with restriction sites that were found among 14 previously sequenced DRB3 alleles. The PCR-RFLP typing method was evaluated on 168 genomic DNA samples collected from animals of 10 cattle breeds, 48 of which were typed in the Fourth International BoLA Workshop for BoLA-DRB and -DQ by conventional restriction fragment length polymorphism (RFLP) analysis using heterologous and homologous DNA probes. Thirty-one DRB/DQ haplotypes containing 23 DRB3 alleles were identified among the 48 workshop animals analysed. Using PCR-RFLP, 11 DRB3 alleles were identified in 18 workshop animals for which DRB RFLPs were not informative. PCR-RFLP typing of additional animals revealed five new DRB3 alleles, of which three contained a putatively located three basepair deletion in the identical position as found for the sequenced allele DRB*2A. PCR-RFLP was shown to be a rapid and sensitive method for the detection of polymorphism in a functionally relevant domain of the BoLA-DRB3 gene and should be useful for studying the evolution of DRB polymorphism in cattle and other Bovidae.

Alleles↗
PubMed1995

Identification and characterization of new BoLA-DRB3 alleles by heteroduplex analysis and direct sequencing.

A sample of 52 mixed-breed dairy cattle (Holstein Friesian and Jersey) and 51 beef cattle (Hereford) from south-east Queensland was studied. The second exon of BoLA-DRB3 was amplified by polymerase chain reaction (PCR), and polymorphisms were detected by heteroduplex analysis. A large number of different heteroduplex patterns indicated extensive sequence polymorphism. Direct sequencing of PCR products from 17 homozygotes and cloning and sequencing of PCR product from two heterozygotes resulted in the identification and characterization of four novel alleles. The previously described allele BoLA-DRB3*2A is characterized by an amino acid deletion at position 65. We have identified three animals that are homozygous for this amino acid deletion, indicating that the deletion is unlikely to result in loss of function. Two of these animals had allele BoLA-DRB3*2A, and one had a novel allele with codon 65 deleted but differing from BoLA-DRB3*2A at a number of other amino acid positions. In conclusion, heteroduplex analysis allows rapid discrimination between homozygotes and heterozygotes, and enables rapid identification of new BoLA-DRB3 alleles.

Alleles↗
PubMed1987

Anti-BoLA-w8 monoclonal antibody: production of a tissue typing reagent after blocking monomorphic sites on bovine mononuclear cells.

A murine monoclonal antibody, WIM-8 with specificity for BoLA-w8, was generated after using an immunization protocol designed to reduce the response to bovine monomorphic determinants. BoLA-w8/w9 peripheral blood mononuclear (PBM) cells were incubated with an antiserum from mice immunized with BoLA-w10/w20 PBM cells. Splenocytes from a mouse that received three intravenous injections of the treated cells were fused with murine myelomas. A hybridoma supernatant with alloreactivity in ELISA against BoLA-w8 positive but not BoLA-w8 negative PBM cells was tested for lymphocytotoxicity in parallel with parous alloantisera. WIM-8, an IgG2b monoclonal antibody, identified the 44,000 dalton protein of the bovine MHC class I molecule present on Bos taurus and Bos indicus animals.

Animals↗
PubMed2005

New polymorphisms for the BoLA-DRB3 upstream regulatory region.

Two new alleles, named BoLA-DRB3-P*06 and BoLA-DRB3-P*07, have been identified for the upstream regulatory region of the BoLA-DRB3 gene. The 228-bp nucleotide sequences of the promoter comprising the W, X, Y, CAAT and TATA regulatory boxes were analysed. The BoLA-DRB3-P*06 exhibits one insertion between the W and X boxes, and one transition between the X and Y boxes. On the other hand, the BoLA-DRB3-P*07 showed one insertion in the X box.

Animals↗
PubMed2004

Effect of Escherichia coli morphogene bolA on biofilms.

Biofilm physiology is established under a low growth rate. The morphogene bolA is mostly expressed under stress conditions or in stationary phase, suggesting that bolA could be implicated in biofilm development. In order to verify this hypothesis, we tested the effect of bolA on biofilm formation. Overexpression of bolA induces biofilm development, while bolA deletion decreases biofilms.

Biofilms↗
PubMed2003

Biological effect of varying peptide binding affinity to the BoLA-DRB3*2703 allele.

MHC class I and II molecules are immunoregulatory cell surface glycoproteins, which selectively bind to and present antigenic peptides to T-lymphocytes. Murine and human studies show that variable peptide binding affinity to MHC II molecules influences Th1/Th2 responses by inducing distinctive cytokine expression. To examine the biological effects of peptide binding affinity to bovine MHC (BoLA), various self peptides (BoLA-DQ and fibrinogen fragments) and non-self peptides from ovalbumin (OVA), as well as VP2 and VP4 peptides from foot and mouth disease virus (FMD-V) were used to (1) determine binding affinities to the BoLA-DRB3*2703 allele, previously associated with mastitis susceptibility and (2) determine whether peptide binding affinity influences T-lymphocyte function. Peptide binding affinity was determined by a competitive assay using high affinity biotinylated self-peptide incubated with purified BoLA-DRB3*2703 in the presence of various concentrations of competing peptides. The concentrations of non-self peptide required to inhibit self-peptide binding by 50% (IC50) were variable, ranging from 26.92 to > 320 microM. Peptide-specific T-lymphocyte function was determined by measuring DNA synthesis, cell division, and IFN-gamma production in cultures of mononuclear cells from a BoLA-DRB3*2703 homozygous cow. When compared to non-stimulated control cultures, differences in lymphocyte function were observed for all of the assessed parameters; however, peptide-binding affinity did not always account for the observed differences in lymphocyte function.

Alleles↗
PubMed1989

Preliminary report on BoLA polymorphism in Guernsey cattle.

A total of 95 registered Guernsey cows and heifers sired by 34 bulls were typed for class I antigens encoded by the bovine major histocompatibility complex (BoLA). A panel of alloantisera was used to detect 21 of the 33 internationally recognized BoLA specificities. Fourteen BoLA specificities were detected in the herd using a standard lymphocyte microcytotoxicity test. The most frequent BoLA specificity detected was w6; but in 85% of these animals, a subtype of w6, either w17 (13.7%) or UR6.2 (34.7%), could be assigned. The next most common BoLA specificities, by frequency, were w12, w15, UR1, and w21. Specificities not detected were w2, w3, w5, w7, w11, w16, w19, w20, and w24.

Animals↗
PubMed2003

[Genetic mechanisms of resistance and susceptibility to leukemia in Ayrshire and black pied cattle breeds determined by allelic distribution of gene Bola-DRB3].

In the herds of Ayrshire and Black Pied cattle breeds of Russian selection, comparative analysis of allelic distribution of BoLA-DRB3 was performed in animal groups with different status of persistent lymphocytosis (PL) caused by the bovine leukemia virus (BLV). Alleles were typed by PCR-RFLP. Different spectra of BoLA-DRB3 alleles mediating susceptibility and resistance to leukemia were detected in the studied breeds. The role of amino acid motives in beta 1 domain of BoLA-DRB3 antigens was confirmed: ER (in positions 70-71), in resistance to leukemia and VDTY and VDTV (75-78), in susceptibility to leukemia. The nucleotide sequence of allele BoLA-DRB3.2*7 with deletion of codon 65, which resulted in the changed conformation of the corresponding antigen molecule, was associated with resistance to PL. Cows of Black Pied and Ayrshire breeds with genotypes coding VDTY/VDTV (RR = 11.67, P = 0.014) and VDTY/VDTY (RR = 4.71, P = 0.022), respectively, were shown to be susceptible to PL. The role of heterozygosity level was demonstrated (estimated by BoLA-DRB3 alleles and by amino acid motives in positions 75-78 of the antigen) as an unspecific factor of resistance to PL. The lowest heterozygosity level by amino acid motives (75-78) was revealed in PL animals, for which sample inbreeding coefficients were detected: F = 0.324 and F = 0.084 in Ayrshire and Black Pied breeds, respectively.

Alleles↗