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PMID:11043467
| Citation |
Day, RB, McAlvin, CB, Loh, JT, Denny, RL, Wood, TC, Young, ND and Stacey, G (2000) Differential expression of two soybean apyrases, one of which is an early nodulin. Mol. Plant Microbe Interact. 13:1053-70 |
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| Abstract |
Two cDNA clones were isolated from soybean (Glycine soja) by polymerase chain reaction with primers designed to conserved motifs found in apyrases (nucleotide phosphohydrolase). The two cDNAs are predicted to encode for two, distinct, apyrase proteins of approximately 50 kDa (i.e., GS50) and 52 kDa (i.e., GS52). Phylogenetic analysis indicated that GS52 is orthologous to a family of apyrases recently suggested to play a role in legume nodulation. GS50 is paralogous to this family and, therefore, likely plays a different physiological role. Consistent with this analysis, GS50 mRNA was detected in root, hypocotyls, flowers, and stems, while GS52 mRNA was found in root and flowers. Neither gene was expressed in leaves or cotyledons. Inoculation of roots with Bradyrhizobium japonicum, nitrogen-fixing symbiont of soybean, resulted in the rapid (<6 h) induction of GS52 mRNA expression. The level of GS50 mRNA expression was not affected by bacterial inoculation. Western blot (immunoblot) analysis of GS50 expression mirrored the results obtained by mRNA analysis. However, in contrast to the mRNA results, GS52 protein was found in stems. Interestingly, anti-GS52 antibody recognized a 50-kDa protein found only in nodule extracts. Treatment of roots with anti-GS52 antibody, but not anti-GS50 antibody or preimmune serum, blocked nodulation by B. japonicum. Fractionation of cellular membranes in sucrose density gradients and subsequent Western analysis of the fractions revealed that GS50 colocalized with marker enzymes for the Golgi, while GS52 colocalized with marker enzymes for the plasma membrane. Restriction fragment length polymorphism (RFLP)-based mapping placed the gs52 gene on major linkage group J of the integrated genetic map of soybean. These data suggest that GS50 is likely an endo-apyrase involved in Golgi function, while GS52 is localized on the root surface and appears to play an important role in nodulation. |
| Links |
PubMed Online version:10.1094/MPMI.2000.13.10.1053 |
| Keywords |
Amino Acid Sequence; Antibodies/immunology; Apyrase/genetics; Apyrase/immunology; Apyrase/isolation & purification; Apyrase/metabolism; Bradyrhizobium/physiology; Chromosome Mapping; DNA Primers; DNA, Complementary; Gene Expression Regulation, Plant; Genes, Plant; Membrane Proteins; Molecular Sequence Data; Nucleic Acid Hybridization; Plant Proteins/genetics; Plant Proteins/metabolism; Plant Roots/enzymology; Plant Roots/microbiology; Plant Structures/enzymology; Polymerase Chain Reaction; RNA, Messenger/genetics; RNA, Messenger/metabolism; RNA, Plant/genetics; RNA, Plant/metabolism; Recombinant Fusion Proteins/biosynthesis; Recombinant Fusion Proteins/isolation & purification; Soybeans/enzymology; Soybeans/genetics; Soybeans/microbiology; Soybeans/physiology |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0005886: plasma membrane |
ECO:0000314: |
C |
Figure 10 shows that GS52 eluted in fractions that had plasma membrane markers. |
complete | ||||
| GO:0009877: nodulation |
ECO:0000314: |
P |
Figure 5B shows upregulation of GS52 in response to the nodule forming B. japonicum, and figure 8 shows that sections of roots treated with anti-GS52 antibody develop fewer B. japonicum nodules. This "strongly argues for the involvement of GS52 as an essential component of the soybean nodulation response." |
complete | ||||
|
part_of |
GO:0005886: plasma membrane |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0000139: Golgi membrane |
ECO:0000314: |
C |
Figure 10 shows GS50 eluted in the fraction with a golgi membrane marker. |
complete | ||||
|
part_of |
GO:0000139: Golgi membrane |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
See also
References
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