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Gene frequencies of DRB3.2 locus of Argentine Creole cattle.

Gene and genotype frequencies of BoLA-DRB3 were studied in seven herds of Argentine Creole cattle. Twenty-one out of thirty previously identified alleles were detected in this breed. The F statistics showed high degree of variability among the studied subpopulations suggesting that subdivision and genetic drift, rather than inbreeding, have been the major factors acting on the observed interpopulation variability. The observed high degree of genetic variation in Argentine Creole cattle could be crucial for the long-term survival of this population. Maintenance of such polymorphism, as a genetic resource, could be an important issue that will demand attention in future breeding programmes in species under high selective pressures.

Alleles↗

Conservation of promoter, coding and intronic regions of the non-classical MHC class II DYA gene suggests evolution under functional constraints.

The major histocompatibility complex (MHC) in ruminants contains a unique pair of class II genes (DYA and DYB) of unknown function. As functional genes show higher levels of nucleotide conservation than pseudogenes we compared the DYA genes from sheep and cattle, two species which diverged from a common ancestor approximately 20 million years ago. Comparative analysis identified levels of nucleotide conservation in immediate promoter (97%), coding (94%) and intronic regions (91%) comparable with functional MHC genes. The Ovar-DYA transcript revealed an open reading frame encoding a 288 amino acid protein compared with a 253 amino acid protein associated with the BoLA-DYA transcript. A dinucleotide deletion in exon 4 of the Ovar-DYA transcript combined with alternative exon 5 splice sites introduces unusual diversity to the cytoplasmic domain of the Ovar-DYalpha polypeptide. The degree of conservation between these class II MHC genes is consistent with evolution under purifying selection suggesting that these genes retain a unique function in ruminants.

Amino Acid Sequence↗

Ecological aspects of cattle tick control in central Zambia.

In ecological studies in central Zambia, both climate and ecotype affected population dynamics of tick species. Below average rainfall for several years caused a suppression in numbers of Rhipicephalus appendiculatus Neumann adults. Reduction in rainfall leading to changes in grazing patterns is thought to have been responsible for an increase in numbers of Amblyomma variegatum Fabricius adults in a grassland habitat. There were reasonable correlations between numbers of each tick species on individual hosts over 1 year old. However, there were no relationships between numbers of ticks and bovine lymphocyte antigens (BoLA).

Animals↗

Sigma S-dependent growth-phase induction of the csgBA promoter in Escherichia coli can be achieved in vivo by sigma 70 in the absence of the nucleoid-associated protein H-NS.

The stationary-phase-specific sigma factor sigma S (RpoS/KatF) is required for Escherichia coli to induce expression of fibronectin-binding curli organelles upon reaching stationary phase. We show that the csgA gene which encodes the curlin subunit protein belongs to a dicistronic operon, csgBA. The transcriptional start site of csgBA was determined and an AT-rich up-stream activating sequence (UAS) required for transcriptional activation was identified. The pcsgBA promoter is not specific for sigma S since the same promoter sequence can be used by E sigma 70 in vivo in a strain lacking nucleoid-associated protein H-NS and sigma S. Transcription remained growth-phase induced and dependent upon the UAS in such a double mutant. Furthermore, we demonstrate that an additional operon, hdeAB, which is also dependent upon sigma S for transcription, can be transcribed by E sigma 70 in vivo in the absence of H-NS by utilizing the phdeAB promoter. Two other genes known to be under the control of sigma S for expression, bolA and katE, remained transcriptionally silent in the absence of H-NS. It is suggested that a subset of E. coli promoters can be recognized by both E sigma S and E sigma 70 in vivo but H-NS interacting with these sequences prevents formation of successful transcription-initiation complexes with E sigma 70.

Amino Acid Sequence↗

Poly(A)-polymerase I links transcription with mRNA degradation via sigmaS proteolysis.

Bacteria rapidly adapt to changes in growth conditions through control of transcription and specific mRNA degradation. Interplay of both mechanisms must exist in order to achieve fine-tuned regulation of gene expression. Transcription of the Escherichia coli bolA gene is mediated by the RpoS/sigmaS transcription factor in response to environmental signals. In this report it is shown that the mechanisms of bolA1p mRNA transcription and degradation are tightly connected at the onset of stationary phase and in response to sudden carbon starvation. In stationary phase, bolA1p mRNA levels were reduced 2.5-fold in a poly(A)-polymerase I (PAPI) mutant, explained by the significant threefold reduction in sigmaS protein levels in the same strain. Furthermore, fusions with the rpoS gene, analysis of the stability of sigmaS and the levels of RssB indicate that the absence of PAPI enhances RssB-mediated sigmaS proteolysis specifically in starved cells. The fact that PAPI induces higher cellular levels of a global regulator is a novel finding of wide biological significance. PAPI could work as a linker between transcription and mRNA degradation with the ultimate goal of adapting and surviving to growth-limiting conditions.

Bacterial Proteins↗

Macrophage-mediated cytostasis and lymphocyte cytotoxicity in cattle immunized with Theileria annulata sporozoites or macroschizont-infected cell lines.

Immunization with either sporozoites or macroschizont-infected cell lines protected calves against challenge with lethal doses of sporozoites of Theileria annulata. Stocks from India, Turkey and Morocco all conferred protective immunity to each other, irrespective of the immunizing regime. Although heterogeneous clinical responses were induced by the two immunizing regimes, both stimulated similar patterns of macrophage cytostasis as expressed as an inhibition of proliferation of macroschizont-infected cell lines. Macrophage cytostasis was detected consistently after immunization and after challenge, arising at the same time as macroschizonts were detectable. Its expression was sustained and inhibited the proliferation of both autologous and allogeneic (BoLA-mismatched) cell lines. In contrast, these two immunizing regimes differed in their ability to stimulate the production of cytotoxic cells. Calves immunized with autologous cell lines or sporozoites developed very transient populations of cytotoxic cells expressing only a low level of specific lysis for autologous infected cells; agglutinating antibodies for immunizing or autologous cell lines were not detected in these calves. Calves immunized with allogeneic cell lines produced cytotoxic cells which were specific only for the immunizing cell lines; these calves also produced antibodies which agglutinated the immunizing cell lines.

Animals↗

The development and specificity of cytotoxic cells in cattle immunized with autologous or allogeneic Theileria annulata-infected lymphoblastoid cell lines.

Two groups of animals were immunized with either 10(6) autologous or 10(6) allogeneic Theileria annulata-infected lymphoblastoid cells cultured in vitro. The development and specificity of cytotoxic cells generated in vivo were measured throughout immunization and challenge using a panel of target cells that were either Theileria-infected or uninfected blast cells of known bovine lymphocyte antigen (BoLA) specificities. After inoculation of the cell lines the two groups showed distinct differences in both their clinical responses and the target specificity of the cytotoxic cells detected. The allogeneic T. annulata cell line recipients showed a very mild clinical response, and on day 9 after inoculation a strong cytotoxic response was detected. The response appeared to be directed against the allogeneic major histocompatibility complex (MHC) antigens of the inoculated cell line in some form of graft rejection response. By day 23 the predominant cytotoxic response was directed against the recipient animals' own cells infected with the parasite. In contrast, the autologous T. annulata cell line recipients showed very severe clinical reactions, and low levels of cytotoxicity were detected. The cytotoxicity was directed against parasite-infected targets but did not appear to be MHC restricted until day 20. Both groups were immune to a heterologous sporozoite challenge that proved lethal to two susceptible control animals, and on day 10 after challenge a peak of cytotoxicity was detected which was directed against the autologous infected target cell. This would suggest that this cytotoxic response was MHC restricted and was also cross-reactive between the heterologous parasite stocks used.

Animals↗

Cross reaction of monoclonal antibodies to human MHC class I and class II products with bovine lymphocyte subpopulations.

Peripheral blood lymphocytes (PBL) from cattle have been separated into T & B cell subpopulations using a panning technique. These T and B cell preparations have then been tested by direct and indirect complement mediated cytotoxicity tests with a series of monoclonal antibodies (Moabs) reacting with HLA class I and class II products. The anti-class I monoclonals were monomorphic or non-reactive and where reactive killed both B and T cells. The anti-class II monoclonals reacted only with B lymphocytes. There was no relationship between their reactivity pattern and any BoLA specificity.

Animals↗

Determinant role of E. coli RNase III in the decay of both specific and heterologous mRNAs.

A comparative analysis of mRNA decay was carried out in Escherichia coli using the wild-type and an isogenic RNase III deletion strain. We have studied the mRNA degradation from the Escherichia coli gene bolA, the Lactococcus lactis biovar diacetylactis citQRP operon and the Desulfovibrio vulgaris Hildenborough gene cyc. As seen by a dramatic stabilization of the specific mRNAs in the mutant strain, RNase III was crucial for the decay process of these three messages. Since RNase III, unlike RNase E, is not essential for bacterial viability we think that there is potential for using RNase III mutant strains to modulate gene expression.

Bacterial Proteins↗

Cattle MHC: evolution in action?

Because major histocompatibility complex (MHC) genes play a major role in the development of acquired immune responses, it is essential to obtain comparative information on their organisation, expression and possible functional dichotomies in different species. In human, three classical, polymorphic class I genes (HLA-A, B- and -C) and four expressed A/B class II gene pairs (HLA-DM, -DP, -DQ and -DR) are each present on all haplotypes. With the exception of the HLA-DRB loci, it has been assumed that a similar rigid organisational situation exists in other mammalian species. However, extensive analysis of the bovine MHC (BoLA) at both the genomic and transcriptional levels has revealed a degree of genetic fluidity not described in other species. None of the four (or more) classical class I genes identified is consistently expressed, and haplotypes differ from one another in both the number and composition of expressed class I genes. Similarly, in the class II region, the number of DQ genes varies between haplotypes in both number and composition. These variations in both class I and II (which appear to reflect differences at the genomic level) are likely to play an important role in cattle immune responses. The observed phenotypic differences in cattle demonstrate very clearly the dynamic nature of the MHC region. This review addresses the functional impact of such variation in different breeds and populations, and its significance in terms of MHC evolution.

Animals↗

Serologically defined (SD) locus in cattle.

Using cytotoxic serums obtained from multiparous cows or by alloimmunization, we have detected 11 lymphocyte antigens controlled by codominant alleles at a serologically defined locus called BoLA-A (bovine lymphocyte antigens). This locus, along with the lymphocyte defined loci previously reported, establishes the existence of a major histocompatibility system of cattle.

Alleles↗

Theileria annulata in CD5(+) macrophages and B1 B cells.

Theileria parasites infect and transform bovine leukocytes. We have analyzed laboratory-established Theileria sp.-infected leukocyte lines and observed that transformed macrophages express CD5. Low-level expression of CD5 by macrophages was further confirmed on three independent Theileria annulata clinical isolates from Tunisia. Interestingly, the fourth CD5(+) clinical isolate (MB2) was morphologically different, expressed surface immunoglobulin M (IgM) and BoLA class II, and had rearranged Ig light-chain genes. To demonstrate that MB2 did indeed contain CD5(+) B cells, individual clonal lines were obtained by limiting dilution, and CD5 expression and Ig gene rearrangement were confirmed. This suggests that in natural infections T. annulata can invade and transform CD5(+) B cells.

Animals↗

Stationary-phase-inducible "gearbox" promoters: differential effects of katF mutations and role of sigma 70.

Many of the changes in gene expression observed when Escherichia coli cells enter stationary phase are regulated at the level of transcription initiation. A group of stationary-phase-inducible promoters, known as "gearbox" promoter, display a characteristic sequence in the -10 region which differs greatly from the consensus sequence for sigma 70-dependent promoters. Here we describe our studies on the gearbox promoters bolAp1 and mcbAp, responsible for the temporally regulated transcription of bolA and the genes involved in the synthesis of the peptide antibiotic microcin B17, respectively. Deletion analysis of mcbAp demonstrated that the stationary-phase-inducible properties of this promoter are found in a DNA fragment extending from -54 to +11 bp, surrounding the transcriptional start site, and are separable from DNA sequences responsible for the OmpR-dependent stimulation of transcription of mcbAp. In vitro transcription studies indicate that the RNA polymerase holoenzyme involved in the transcription of mcbAp contains sigma 70. In this and an accompanying paper (R. Lange and R. Hengge-Aronis, J. Bacteriol. 173: 4474-4481, 1991), experiments are described which show that the product of katF, a global regulator of stationary-phase gene expression and a putative sigma factor, is required for the expression of bolAp1 fused to the reporter gene lacZ. In contrast, mcbAp appears to be negatively regulated by katF. We discuss the implications of these results for postexponential gene expression and the role of gearbox sequences in the regulation of promoter activity.

Base Sequence↗

Osmotic regulation of rpoS-dependent genes in Escherichia coli.

The rpoS gene, which encodes a putative alternative sigma factor (sigma S), is essential for the expression of a variety of stationary-phase-induced genes as well as for stationary-phase-specific multiple-stress resistance. As previously shown for the otsA and otsB genes (R. Hengge-Aronis, W. Klein, R. Lange, M. Rimmele, and W. Boos, J. Bacteriol. 173:7918-7924, 1991), we demonstrate here that additional rpoS-controlled genes (bolA, csi-5) as well as at least 18 proteins on two-dimensional O'Farrell gels could be induced in growing cells by osmotic upshift via an rpoS-dependent mechanism. Also, rpoS-dependent thermotolerance and resistance against hydrogen peroxide could be osmotically stimulated. In contrast, the expression of glgS, while exhibiting strong stationary-phase induction, was only weakly increased by elevated osmolarity, and several rpoS-dependent proteins previously identified on two-dimensional gels were not osmotically induced. During osmotic induction of rpoS-dependent genes, rpoS transcription and the level of sigma S remained unchanged. We conclude that osmotically regulated genes represent a subfamily within the rpoS regulon that requires differential regulation in addition to that provided by sigma S.

Bacterial Proteins↗

Chromosome banding and gene localizations support extensive conservation of chromosome structure between cattle and sheep.

By using three gene probes, one derived from the porcine major histocompatibility complex (MHC) and two from bovine cytokeratin genes, type I (KRTA) and type II (KRTB), the hypothesis of conservation of genome structure in two members of the family Bovidae was examined. Gene mapping data revealed the MHC to be in chromosome region 23q15----q23 in cattle (BOLA) and 20q15----q23 in sheep (OLA). KRTA was localized to chromosome region 19q25----q29 in cattle and 11q25----q29 in sheep and KRTB to 5q14----q22 in cattle and 3q14----q22 in sheep. The banding patterns of the chromosome arms to which the loci were assigned were identical in both species. Moreover, the resemblances of GTG- or QFQ-banding patterns between the cattle and sheep karyotypes illustrated further chromosome homologies. These studies, based on gene mapping comparisons and comparative cytogenetics, document that within bovid chromosomes, homology of banding patterns corresponds to a homologous genetic structure. Hence, we propose that gene assignments on identified chromosomal segments in one species of the Bovidae can be extrapolated, in general, to other bovid species based on the banding homologies presented here.

Animals↗

Renitelo cattle dermatophilosis and PCR-RFLP analysis of MHC gene.

Renitelo breed is a cattle breed created at Kianjasoa station (Madagascar) by a triple crossing Malagasy Zebu x Limousine x Afrikander. This breed besides many valuable advantages, such as rapid growth and drought power, presents a huge disadvantage which is sensitivity to skin disease, dermatophilosis, previously known as streptotrichosis. This disease caused by Dermatophilus congolensis is one of the major threats for the population of Renitelo cattle. An allele of MHC gene has been shown to be dramatically associated to hypersensitivity to the disease in other cattle breed. To bring further information to tick borne disease clinical survey, mainly dermatophilosis, we wanted to verify if such allele could be found in this breed. Renitelo cattle included in this study were chosen for the presence of dermatophilosis lesions in more or less severe form (N = 17). These animals were blood sampled and a genetic analysis on the MHC gene BoLA-DRB3 was performed, by PCR amplification using BOD 31 & BOD 32 primers. Amplified products were analyzed by RFLP using enzymes. Restriction band profiles were characterized according to previously defined patterns. Three cows out of the 17 cattle analyzed for MHC gene presented the hypersensitive allele FDA. Two out of the three hypersensitive cows were pure breed while one was half breed. All the cows presented dermatophilosis lesions at least during rainy season but one of them particularly suffered from severe lesions covering all its body and died of the illness. This study shows that hypersensitivity allele found in other bovine breeds can be found in Renitelo breed. This result seemed to suggest that this characterization could be utilized in breeding program for this breed.

Actinomycetales Infections↗

The major histocompatibility complex of ruminants.

Studies of the major histocompatibility complex (MHC) of cattle over the past twenty years have revealed a reasonably detailed picture of the genetic organisation and function of the genes within this genetic system. Serological and biochemical analysis of lymphocyte cell surface antigens provided the first evidence for highly polymorphic MHC genes in cattle and other ruminant species. The MHC of cattle was thus named the bovine leucocyte antigen (BoLA) system. During the past 10 years, tools of molecular biology have been used to characterise the number of MHC genes, their sequence and fine structure in a number of ruminant species. Although individual MHC genes were found to have clear orthologues among ruminants and other mammalian species, the MHC of cattle, and probably that of sheep and goats, has a unique genetic organisation. Cattle have a class II gene cluster (class IIb region) which is physically distant from all the other MHC genes on the same chromosome. Moreover, genes involved in antigen processing, such as the proteosome subunit locus LMP2, are also found in the class IIb region, demonstrating that these genes need not be in close proximity to other MHC genes to function normally. The MHC class I and class II gene products of ruminants present processed peptides to T lymphocytes which mediate helper and cytotoxic functions. Identification of peptide binding motifs of cattle MHC class I molecules indicates that ruminant MHC molecules function in a similar manner to those of mice and humans. These functional studies provide a firm molecular basis for a number of well-documented associations with infectious diseases, although a detailed understanding of the immunogenetic mechanisms underlying these associations has yet to be elucidated.

Animals↗

Class I alleles of the bovine major histocompatibility system and their association with economic traits.

A total of 179 Holstein cows from the Agriculture Canada Research Branch herd at Ottawa and 271 progeny-tested Canadian Holstein bulls were typed for 37 lymphocyte antigens. Each antigen appears to be controlled by a distinct, codominant allele of the bovine lymphocyte antigen (BoLA)-A locus, which is a class I locus of the bovine major histocompatibility system. Only 10 and 16 alleles were present in Holstein cows and bulls, respectively. The association between bovine lymphocyte antigens and economically important traits was examined in gene substitution models. Separate models were used for the cow and bull data. The substitution of the W6.1 allele for the W10 allele was associated with increased protein yield in the milk in both analyses. The results were also consistent with a previous report that the presence of the W10 allele was associated with increased fat percentage compared with some, but not all, of the other alleles. However, more research is necessary to confirm these findings and to determine the biological mechanisms underlying these associations.

Age Factors↗