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Query: UNIPROT:P06889 (Mol)
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Genetic and molecular analysis of the self-incompatibility locus (S-locus) of the crucifer Brassica has led to the characterization of a multigene family involved in pollen-stigma interactions. While the crucifer Arabidopsis thaliana does not have a self-incompatibility system, S-related sequence were detected in this species by cross-hybridization with Brassica DNA probes. In this paper, we show that an A. thaliana S-related sequence, designated AtS1, is expressed specifically in flower buds. Sequence analysis suggests that AtS1 encodes a secreted glycoprotein that is most similar to the Brassica S-locus related protein SLR1. As has been proposed for SLR1, this gene may be involved in determining some fundamental aspect of pollen-stigma interactions during pollination. The molecular and genetic advantages of the Arabidopsis system will provide many avenues for testing this hypothesis.
Mol Gen Genet 1992 Feb
PMID:Structure and expression of AtS1, an Arabidopsis thaliana gene homologous to the S-locus related genes of Brassica. 137 78

A self-incompatible canola-quality Brassica napus ssp. oleifera line (W1) was generated by introgressing the S-locus from a self-incompatible B. campestris plant into the Westar cultivar. Using the polymerase chain reaction (PCR) with primers derived from conserved regions in S-locus glycoprotein (SLG) alleles, the central region of the active SLG gene (910) was obtained. The remaining portions of the cDNA for this 910 gene were subsequently cloned using the PCR-rapid amplification of cDNA ends (RACE) procedure. Sequence analysis revealed that the 910 cDNA show a high degree of sequence similarity to SLG alleles associated with Class I self-incompatible lines. The 910 gene was found to be absent in the original self-compatible cv. Westar (B. napus) and segregated with self-incompatibility in a mixed population generated from a cross between self-incompatible W1 and self-compatible Westar. RNA blot analysis indicated that high levels of 910 mRNAs were present in the stigma as buds approached anthesis. Thus, the SLG allele of W1 transferred from B. campestris via backcrosses to a line of cv. Westar has been identified.
Mol Gen Genet 1992 Aug
PMID:Use of the polymerase chain reaction to isolate an S-locus glycoprotein cDNA introgressed from Brassica campestris into B. napus ssp. oleifera. 150 46

The sequence is reported of a cDNA molecule homologous to an mRNA from stigma tissue of Brassica oleracea plants homozygous for the S5 self-incompatibility allele. This cDNA is closely related to a previously published sequence designated SLR2, which was obtained from the same stigma cDNA library and is also related to the SLR1 gene, a cDNA for which has also been obtained from this library. Various B. oleracea lines differing in S alleles and of different varieties have been screened for the presence of particular S gene family sequences using a method involving hybridization of sequence-specific oligonucleotide probes to PCR products. The results indicate that a gene homologous to the sequence presented here is absent from a line lacking the S5 allele, though present in other lines containing the S5 allele, regardless of their genetic background. This finding suggests that the sequence represents a transcript of the SLG (S locus glycoprotein) gene. A similar approach has confirmed that a cDNA derived from a Brassica line containing the S29 allele is also S allele-specific. The predicted amino acid sequences derived from a number of S gene family sequences are compared using numerical methods and possible evolutionary relationships between them are discussed.
Mol Gen Genet 1992 Mar
PMID:An S5 self-incompatibility allele-specific cDNA sequence from Brassica oleracea shows high homology to the SLR2 gene. 155 30

Three pathogenesis-related (PR) proteins of tobacco are acidic isoforms of beta-1,3-glucanase (PR-2a, -2b, -2c). We have cloned and sequenced a partial cDNA clone (lambda FJ1) corresponding to one of the PR-2 beta-1,3-glucanases. A small gene family encodes the PR-2 proteins in tobacco, and similar genes are present in a number of plant species. We analyzed the stress and developmental regulation of the tobacco PR-2 beta-1,3-glucanases by using northern and western analyses and a new technique to assay enzymatic activity. Stress caused by both thiamine and tobacco mosaic virus (TMV) infection resulted in a dramatic increase in the levels of PR-2 mRNA, protein, and enzyme activities. The increased PR-2 gene expression in upper uninoculated leaves of plants infected with TMV also suggests a role in systemic acquired resistance. During floral development, a number of beta-1,3-glucanase activities were observed in all flower tissues. However, PR-2 polypeptides were observed only in sepal tissue. In contrast, an mRNA that hybridized to the PR-2 cDNA was present in stigma/style tissue and the sepals. Primer extension analysis confirmed the identity of the PR-2 mRNA in sepals, but indicated that the beta-1,3-glucanase gene expressed in the stigma/style of flowers was distinct from the PR-2 genes. The induction of PR-2 protein synthesis by both stress and developmental signals was accompanied by a corresponding increase in the steady-state levels of PR-2 mRNA, suggesting that PR-2 gene expression is regulated, in part, at the level of mRNA accumulation.
Mol Plant Microbe Interact
PMID:Pathogenesis-related acidic beta-1,3-glucanase genes of tobacco are regulated by both stress and developmental signals. 193 13

Self-incompatibility in Brassica oleracea is controlled by a single genetic locus (the S locus) with nearly 50 different alleles. In this paper, we report the characterization of the S2 allele, a pollen recessive self-incompatibility allele that exhibits weak DNA homology to the other previously sequenced S locus glycoprotein genes (SLG-6, -13, -14, -22 from alleles S6, S13, S14 and S22, respectively). Stigma cDNA clones with sequence homology to SLG-13 were isolated from two different S2 homozygous strains belonging to two different B. oleracea cultivars, var. alboglabra (Chinese kale) and var. italica (broccoli). The two S2 cDNA sequences are 90% homologous to each other, but only 70% homologous to SLG-13. Using the Chinese kale S2 genetic background, we demonstrate that the isolated alboglabra cDNA sequence is a transcript from a gene, designated SLG-2A that resides at the S locus, and propose that it is a putative determinant of S2 allelic specificity. Among the estimated 10-15 genomic copies of SLG-related genes detected in the S2 genome, we cloned and characterized the SLG-2A gene and another closely related and genetically linked gene copy, SLG-2B. A complete open reading frame that is 94% homologous to SLG-2A is located within SLG-2B. The existence of this intact duplicated S gene raises the possibility that SLG-2B may also be involved in the functioning of self-incompatibility in Brassica oleracea.
Mol Gen Genet 1990 Jul
PMID:A new class of S sequences defined by a pollen recessive self-incompatibility allele of Brassica oleracea. 198 Mar 34

The sporophytic self-incompatibility system of Brassica species is controlled by a single locus, S. Recognition of self between pollen and stigma is probably mediated by S locus-specific glycoproteins (SLSGs). We describe the isolation, from an S29 homozygote of Brassica oleracea, of two different cDNA clones for transcripts which are equally abundant in stigmas competent for self-incompatibility and each of which is homologous to previously reported SLSG sequences. Extensive DNA sequence divergence between the two clones precludes their cross-hybridisation and each acts as a gene-specific probe. All S genotypes appear to have a single copy of each gene but there are significantly different levels of polymorphism associated with each. The clear structural homology between the two indicates a gene duplication involving the S locus and, perhaps, related to the evolution of self-incompatibility.
Mol Gen Genet 1989 Jul
PMID:A homozygous S genotype of Brassica oleracea expresses two S-like genes. 255 Jul 59

The three-dimensional structures of a series of 6-kDa trypsin inhibitors isolated from the stigma of the ornamental tobacco Nicotiana alata have been determined by 1H NMR spectroscopy combined with simulated annealing calculations. The proteins, T1-T4, are proteolytically cleaved from a 40.3-kDa precursor protein, NA-proPI, together with a chymotrypsin inhibitor, C1, the structure of which was reported recently [Nielsen, K.J., Heath, R.L., Anderson, M.A., & Craik, D.J. (1994) J. Mol. Biol. 242, 231-243]. Each of the proteinase inhibitors comprises 53 amino acids, including 8 cysteine residues which are linked to form 4 disulfide bridges. The proteins have a high degree of sequence identity and differ mainly in residues around the putative reactive sites. The structure of T1 was determined using a set of 533 interproton distance restraints derived from NOESY spectra, combined with 33 dihedral restraints derived from 3JNH-H alpha coupling constants and 16 hydrogen bonds. The structures of the remaining inhibitors (T2-T4) were deduced to be almost identical to T1, on the basis of their similar chemical shifts and 2D spectra. The current study demonstrates that the structures of the trypsin inhibitors (T1-T4) are similar to that previously found for the chymotrypsin inhibitor, C1. Despite differences in sequence, there is conservation in backbone geometry between the reactive site loops of the two classes of inhibitors. From this, it is clear that the nature of the side chain on the primary binding residue, rather than the backbone fold, is the main determinant of the enzyme specificities of these proteinase inhibitors.
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PMID:Structures of a series of 6-kDa trypsin inhibitors isolated from the stigma of Nicotiana alata. 757 34

A partial cDNA clone, Pis 63, corresponding to a mRNA highly expressed in Brassica napus pistils, was isolated by differential screening. PCR was used to complete the Pis 63 sequence (Pis 63-1) and to obtain the sequence of another related cDNA (Pis 63-2). Northern blot and in situ analyses demonstrated that these transcripts are expressed in the stigma throughout flower development. Pis 63-1 and Pis 63-2 display similarity to a cotton fibre cDNA clone.
Plant Mol Biol 1994 Nov
PMID:Molecular analysis of two Brassica napus genes expressed in the stigma. 781 80

Deletions were made in the promoter of the acyl carrier protein (ACP) Acll.2 gene from Arabidopsis to investigate the nature of the cis-acting elements that direct its expression. These constructs, which included the untranslated leader region, were fused to a reporter gene coding for beta-glucuronidase (GUS) and transformed into tobacco. Quantitative fluorometric analysis of GUS activity in transgenic plants showed that expression in young leaves drops to a basal level when a 85 bp domain, from -320 to -236 relative to transcription initiation, is deleted. Maximum promoter activity in roots also depends on this domain, but two other regions are also important. In total, deletion of the sequences from -466 to -55 caused an ca. 80-fold reduction in Acl1.2 promoter activity in roots. The -320 to -236 domain forms a complex with a protein factor found in leaves and roots, which was not detectable in seeds. The formation of this protein-DNA complex was abolished by mutation of a bZIP core motif, ACGT, found within the context AAGACGTAG, which is dissimilar to the other bZIP-binding sites thus far characterized in plants. Previously we showed that Acl1.2 promoter activity is highest in seeds [2]. Here we find, in contrast to leaves and roots, that deletion to position -236 has no effect on GUS levels in seeds. However, nearly a 100-fold drop was observed when the -235 to -55 region was removed. Hence, this 180 bp domain contains all the cis-acting information necessary for Acl1.2 promoter activity in seeds. The same region is necessary for Acl1.2 activity in the receptacle, stigma, tapetum and pollen of the flower, as demonstrated by histochemical staining.
Plant Mol Biol 1994 Dec
PMID:Identification of domains in an Arabidopsis acyl carrier protein gene promoter required for maximal organ-specific expression. 785 29

The three-dimensional structure and disulfide connectivities of a 6-kDa protein isolated from the stigma of the ornamental tobacco Nicotiana alata has been determined by 1H NMR spectroscopy combined with simulated annealing calculations. The protein, termed C1, is a chymotrypsin inhibitor and is one of five homologous proteinase inhibitors that are proteolytically cleaved from a 40.3-kDa precursor protein. The other four proteinase inhibitors (T1 to T4) contain reactive sites for trypsin. The three-dimensional structure of C1 is generally well defined and contains a triple stranded beta-sheet as the dominant secondary structural feature. Several turns and a short region of 3(10) helix are also present. The putative chymotrypsin reactive site is present on an exposed loop which is less defined than the rest of the protein. The overall shape of C1 is disc-like and the N and C termini are exposed, supporting the proposal that this protein results from post-translational processing of the 40.3-kDa precursor protein.
J Mol Biol 1994 Sep 23
PMID:The three-dimensional solution structure by 1H NMR of a 6-kDa proteinase inhibitor isolated from the stigma of Nicotiana alata. 808 44


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