The MADS-domain protein AGAMOUS-like 15 accumulates in embryonic tissues with diverse origins. (9/1068)

AGL15 (AGAMOUS-like 15), a member of the MADS-domain family of regulatory factors, accumulates preferentially in the organs and tissues derived from double fertilization in flowering plants (i.e. the embryo, suspensor, and endosperm). The developmental role of AGL15 is still undefined. If it is involved in embryogenesis rather than some other aspect of seed biology, then AGL15 protein should accumulate whenever development proceeds in the embryonic mode, regardless of the origin of those embryos or their developmental context. To test this, we used AGL15-specific antibodies to analyze apomictic embryogenesis in dandelion (Taraxacum officinale), microspore embryogenesis in oilseed rape (Brassica napus), and somatic embryogenesis in alfalfa (Medicago sativa). In every case, AGL15 accumulated to relatively high levels in the nuclei of the embryos. AGL15 also accumulated in cotyledon-like organs produced by the xtc2 (extra cotyledon2) mutant of Arabidopsis and during precocious germination in oilseed rape. Furthermore, the subcellular localization of AGL15 appeared to be developmentally regulated in all embryogenic situations. AGL15 was initially present in the cytoplasm of cells and became nuclear localized before or soon after embryogenic cell divisions began. These results support the hypothesis that AGL15 participates in the regulation of programs active during the early stages of embryo development.  (+info)

The S15 self-incompatibility haplotype in Brassica oleracea includes three S gene family members expressed in stigmas. (10/1068)

Self-incompatibility in Brassica is controlled by a single, highly polymorphic locus that extends over several hundred kilobases and includes several expressed genes. Two stigma proteins, the S locus receptor kinase (SRK) and the S locus glycoprotein (SLG), are encoded by genes located at the S locus and are thought to be involved in the recognition of self-pollen by the stigma. We report here that two different SLG genes, SLGA and SLGB, are located at the S locus in the class II, pollen-recessive S15 haplotype. Both genes are interrupted by a single intron; however, SLGA encodes both soluble and membrane-anchored forms of SLG, whereas SLGB encodes only soluble SLG proteins. Thus, including SRK, the S locus in the S15 haplotype contains at least three members of the S gene family. The protein products of these three genes have been characterized, and each SLG glycoform was assigned to an SLG gene. Evidence is presented that the S2 and S5 haplotypes carry only one or the other of the SLG genes, indicating either that they are redundant or that they are not required for the self-incompatibility response.  (+info)

Comparison of phylloquinone bioavailability from food sources or a supplement in human subjects. (11/1068)

Phylloquinone (K) absorption was assessed in 22- to 30-y-old human subjects consuming a standard test meal [402 kcal (1682 kJ), 27% energy from fat]. The absorption of phylloquinone, measured over a 9-h period as the area under the curve (AUC), was higher (P < 0.01) after the consumption of a 500- microgram phylloquinone tablet [27.55 +/- 10.08 nmol/(L. h), n = 8] than after the ingestion of 495 microgram phylloquinone as 150 g of raw spinach [4.79 +/- 1.11 nmol/(L. h), n = 3]. Less phylloquinone (P < 0.05) was absorbed from 50 g of spinach (AUC = 2.49 +/- 1.11 nmol/(L. h) than from 150 g of spinach. Absorption of phylloquinone from fresh spinach (165 microgram K), fresh broccoli (184 microgram K) and fresh romaine lettuce (179 microgram K) did not differ. There was no difference in phylloquinone absorption from fresh or cooked broccoli or from fresh romaine lettuce consumed with a meal containing 30 or 45% energy as fat.  (+info)

Molybdenum absorption and utilization in humans from soy and kale intrinsically labeled with stable isotopes of molybdenum. (12/1068)

BACKGROUND: Stable-isotope studies of molybdenum metabolism have been conducted in which molybdenum was added to the diet and was assumed to be absorbed and utilized similarly to the molybdenum in foods. OBJECTIVE: Our objective was to establish whether the molybdenum in foods is metabolized similarly to molybdenum added to the diet. DESIGN: We first studied whether sufficient amounts of molybdenum stable isotopes could be incorporated into wheat, kale, and soy for use in a human study. Enough molybdenum could be incorporated into soy and kale to study molybdenum absorption and excretion. Two studies were then conducted, one in women and one in men. In the first study, each meal contained approximately 100 microg Mo from soy, kale, and extrinsic molybdenum. In the second study, soy and extrinsic molybdenum were compared; the meal contained approximately 300 microg Mo. RESULTS: In the first study, molybdenum was absorbed equally well from kale and an extrinsic source. However, the molybdenum in soy was less well absorbed than the molybdenum in kale or that added to the diet. In the second study, absorption of molybdenum from soy was less than from the extrinsic label. Urinary excretion of soy molybdenum was also lower than urinary excretion of the extrinsic label, but excretion as a percentage of the absorbed dose was not significantly different between treatments. CONCLUSIONS: The molybdenum in soy is less available than molybdenum added to the diet, but the molybdenum in kale is as available as molybdenum added to the diet. Once absorbed, excretion is not significantly different for soy, kale, and extrinsic molybdenum.  (+info)

Mutational analyses of the putative calcium binding site and hinge of the turnip crinkle virus coat protein. (13/1068)

The turnip crinkle carmovirus (TCV) coat protein (CP) is folded into R (RNA-binding), S (shell), and P (protruding) domains. The S domain is an eight-stranded beta barrel common to the coat protein subunits of most RNA viruses. A five-amino-acid hinge connects the S and P domains. In assembled particles, each pair of CP subunits is thought to bind a single calcium ion through interactions with three residues of one subunit and two residues of a neighboring subunit. These five residues comprise the putative calcium-binding site (CBS). The putative CBS and hinge are adjacent to one another. Mutations were introduced into the putative CBS or hinge in an effort to further determine the biological functions of TCV CP. One putative CBS mutant, TCV-M32, exhibited wild-type cell-to-cell movement but failed to move systemically in Nicotiana benthamiana, and particles were not detected. Another putative CBS mutant, TCV-M23, exhibited deficient cell-to-cell movement but particles accumulated in isolated protoplasts. Two other putative CBS mutants, TCV-M22 and -M33, showed wild-type cell-to-cell and systemic movement but elicited mild systemic symptoms that were somewhat delayed. All of the hinge mutants exhibited wild-type movement but some elicited non-wild-type symptoms. Point mutations in the putative CBS or hinge appear to alter virus-ion interactions, secondary structure, or particle conformation, thereby affecting interactions between the CP and plant hosts.  (+info)

An expressional system of human cytochrome P-450 CYP1A1 gene transcription. (14/1068)

AIM: To explore an expressional system of human cytochrome P-450 CYP1A1 (CYP1A1) gene transcription. METHODS: The plasmid pMC 6.3 K containing human CYP1A1 promoter was transiently transfected into Hep G2 cells. The expression of chloramphenical acetyltransferase (CAT) reporter gene was detected by ELISA. RESULTS: Both the CAT expression and CYP1A1 activity increased with the concentrations of beta-naphthoflavone from 2.5 to 10 mumol.L-1. At 10 mumol.L-1 of beta-naphthoflavone, the levels of CAT and CYP1A1 were 94-fold and 2.8-fold those of the corresponding control, respectively. Using this method, the study of 8 glucosinolates with various side chains on the induction of CYP1A1 gene transcription showed that none of the parent glucosinolates increased CAT expression, whereas the breakdown products of indol-3-yl-methyl glucosinolate (glucobrassicin), rather than indole-3-carbinol, increased the CAT expression. CONCLUSION: The CYP1A1 gene transcriptional system was more reliable and sensitive.  (+info)

Glyoxalase I from Brassica juncea is a calmodulin stimulated protein. (15/1068)

Brassica juncea glyoxalase I (S-lactoylglutathione-lyase, EC 4.4.1. 5) is a 56 kDa, heterodimeric protein. It requires magnesium (Mg2+) for its optimal activity. In this report we provide biochemical evidence for modulation of glyoxalase I activity by calcium/calmodulin (Ca2+/CaM). In the presence of Ca2+ glyoxalase I showed a significant (2.6-fold) increase in its activity. It also showed a Ca2+ dependent mobility shift on denaturing gels. Its Ca2+ binding was confirmed by Chelex-100 assay and gel overlays using 45CaCl2. Glyoxalase I was activated by over 7-fold in the presence of Ca2+ (25 microM) and CaM (145 nM) and this stimulation was blocked by the CaM antibodies and a CaM inhibitor, trifluroperazine (150 microM). Glyoxalase I binds to a CaM-Sepharose column and was eluted by EGTA. The eluted protein fractions also showed stimulation by CaM. The stimulation of glyoxalase I activity by CaM was maximum in the presence of Mg2+ and Ca2+; however, magnesium alone also showed glyoxalase I activation by CaM.  (+info)

Characterization and immunolocalization of a cytosolic calcium-binding protein from Brassica napus and Arabidopsis pollen. (16/1068)

Two low-molecular-weight proteins have been purified from Brassica napus pollen and a gene corresponding to one of them has been isolated. The gene encodes an 8.6-kD protein with two EF-hand calcium-binding motifs and is a member of a small gene family in B. napus. The protein is part of a family of pollen allergens recently identified in several evolutionarily distant dicot and monocot plants. Homologs have been detected in Arabidopsis, from which one gene has been cloned in this study, and in snapdragon (Antirrhinum majus), but not in tobacco (Nicotiana tabacum). Expression of the gene in B. napus was limited to male tissues and occurred during the pollen-maturation phase of anther development. Both the B. napus and Arabidopsis proteins interact with calcium, and the potential for a calcium-dependent conformational change was demonstrated. Given this affinity for calcium, the cloned genes were termed BPC1 and APC1 (B. napus and Arabidopsis pollen calcium-binding protein 1, respectively). Immunolocalization studies demonstrated that BPC1 is found in the cytosol of mature pollen. However, upon pollen hydration and germination, there is some apparent leakage of the protein to the pollen wall. BPC1 is also concentrated on or near the surface of the elongating pollen tube. The essential nature of calcium in pollen physiology, combined with the properties of BPC1 and its high evolutionary conservation suggests that this protein plays an important role in pollination by functioning as a calcium-sensitive signal molecule.  (+info)