Leghemoglobin biosynthesis in soybean root nodules. Characterization of the nascent and released peptides and the relative rate of synthesis of the major leghemoglobins

Verma, Desh Pal S.; Ball, Stephan; Guérin, Claude; Wanamaker, Laura

doi:10.1021/bi00570a016

Cited by 72 publications

(21 citation statements)

References 45 publications

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“…1C), which is consistent with our earlier observation (18) as confirmed recently (33) in pea nodules. Thus, subcellular locations of these two nodule-specific proteins are in agreement with all biochemical data (6,10,18,34 Fig. 4).…”

Section: Resultssupporting

confidence: 87%

Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules

Nguyen

Zelechowska

Foster

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

163

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, a subunit of nodule-specific uricase (uricase II) of soybean (Glycine max), is shown to be preferentially synthesized on free polysomes during nodule development and is localized in peroxisomes of the uninfected cells of this tissue. A cDNA clone, isolated by using mRNA from immunoprecipitated polysomes, revealed the primary structure of this protein with a molecular mass of 35,100. That this clone represents N-35 was confirmed by comparing the deduced amino acid sequence with the partial sequence of a CNBr-cleaved peptide of purified N-35. Southern blot hybridizations with genomic DNA suggest that there are several EcoRI fragments containing N-35 sequences. Three of these sequences were isolated from a genomic library of soybean. Nucleotide sequence analysis showed that the complete gene extends almost 5000 base pairs on two EcoRI fragments and the coding region (309 codons) is interrupted by seven introns ranging in size from 154 to 1341 base pairs. Lack of a signal sequence and its translation on free polysomes suggest that N-35 is posttranslationally transported to the peroxisomes. Furthermore, there is no cross-hybridization of N-35 cDNA with RNA from young (3-to 4-day) roots and leaves, indicating that the observed "uricase" activity in these tissues is due to the product of a different gene.Tropical legumes primarily transport fixed nitrogen as ureides, allantoin, and allantoic acid (refs. 1 and 2, see also ref.3). These compounds are formed in root nodules via de novo purine biosynthesis and oxidation. It has been suggested that the enzymes involved in purine biosynthesis are located in the proplastids (4), whereas the oxidation ofpurine occurs in microbodies (peroxisomes) (5-7). A number of enzymes involved in ureide metabolism have been found to be associated with peroxisomes (see refs. 7 and 8), which are more abundant in the uninfected cells of nodules (9). Recent clone from soybean and determined their structures at the nucleotide level. Of general interest are features that may be responsible for the specificity to peroxisomes of this gene product and those involved in tissue-specific induction following infection of a legume plant by Rhizobium. To our knowledge, isolation of a eukaryotic gene that codes for a peroxisomal protein has not been reported previously, and studies of its structure may be of significance in understanding the biogenesis and specialization of the peroxisomes (12, 13) for various metabolic reactions (7,8). Furthermore, comparison of its structure with those of other nodulins (14, 15) should allow identification of sequences that may be responsible for the temporal control in regulating the expression of this gene. MATERIALS AND METHODSPlant Tissues and Isolation of RNA and DNA. Soybean (Glycine max cv. Prize) seeds were inoculated with Rhizobium japonicum (strain 61A76) and germinated as described (16). Infection zone from the primary root of less than a week and nodules from 2-to 3-week-old plants were obtained. Fresh tissues were used for immunocytochemical stud...

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Section: Resultssupporting

confidence: 87%

Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules

Nguyen

Zelechowska

Foster

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

163

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“…Temporally, expression takes place considerably later in the Nif than in the Nod pattern, as has long been known for the genes encoding nitrogenase (Bergersen and Goodchild 1973;Long 1989). Spatially, expression in the Nil pattern, unlike the Nod pattern, is completely absent from the meristematic zone but is most prominent just behind the meristem, like expression of leghemoglobin (Bergersen and Goodchild 1973;Verma et al 1979), upon which nitrogenase function depends. Although expression of Nif pattern genes occurs later and is known to be regulated (by FixLJ-NifA), it does not seem to depend on continued expression of Nod pattern genes per se, as it continues well after nod expression has disappeared.…”

Section: Nil Patternmentioning

confidence: 97%

Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA.

Sharma

Signer²

1990

Genes Dev.

186

106

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Tn5-gusA promoter/probe transposons have been constructed that fuse the Escherichia coli gusA reporter gene transcriptionally or translationally with a target promoter. These have been used to monitor expression of P, hizobium meliloti symbiotic genes within alfalfa nodules. Fusions in all 11 nod genes studied show the same pattern of expression: first on the root surface, then throughout the developing nodule, then mainly in the nodule meristem, falling off progressively through the central region, and then disappearing. In contrast, fusions in all five nif genes studied, all four fix genes, and syrM show a second, different pattern: expression beginning later, first throughout the nodule except for the meristem, strongest just behind the meristem, and falling off progressively through the central region. Novel features revealed by these studies include nod expression in the meristem, regulated in planta expression of control genes nodD1 and nodD3, disappearance of nod expression late in organogenesis, and properties of syrM.

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“…a pink-red color in the infected zone of the nodule was used as a marker of nodule maturity; leghemoglobin gene expression increases proportionally with the age of the nodule [Marcker et al, 1984], and its induction occurs before the detection of nitrogenase activity [Verma et al, 1979] ] mature nodules with leghemoglobin). For each category, nodule numbers were counted with a Leica MZFLIII stereomicroscope equipped with epifluorescence excitation and a GFP longpass filter (500 nm long pass; no.…”

Section: Plant Transformation and Nodule Observationmentioning

confidence: 99%

Large-Scale Analysis of Putative Soybean Regulatory Gene Expression Identifies a Myb Gene Involved in Soybean Nodule Development

et al. 2009

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Nodulation is the result of a symbiosis between legumes and rhizobial bacteria in soil. This symbiosis is mutually beneficial, with the bacteria providing a source of nitrogen to the host while the plant supplies carbon to the symbiont. Nodule development is a complex process that is tightly regulated in the host plant cell through networks of gene expression. In order to examine this regulation in detail, a library of quantitative reverse transcription-polymerase chain reaction primer sets was developed for a large number of soybean (Glycine max) putative regulatory genes available in the current expressed sequence tag collection. This library contained primers specific to soybean transcription factor genes as well as genes involved in chromatin modification and translational regulation. Using this library, we analyzed the expression of this gene set during nodule development. A large number of genes were found to be differentially expressed, especially at the later stages of nodule development when active nitrogen fixation was occurring. Expression of these putative regulatory genes was also analyzed in response to the addition of nitrate as a nitrogen source. This comparative analysis identified genes that may be specifically involved in nitrogen assimilation, metabolism, and the maintenance of active nodules. To address this possibility, the expression of one such candidate was studied in more detail by expressing in soybean roots promoter β-glucuronidase and green fluorescent protein fusions. This gene, named Control of Nodule Development (CND), encoded a Myb transcription factor gene. When the CND gene was silenced, nodulation was reduced. These results, associated with a strong expression of the CND gene in the vascular tissues, suggest a role for CND in controlling soybean nodulation.

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Leghemoglobin biosynthesis in soybean root nodules. Characterization of the nascent and released peptides and the relative rate of synthesis of the major leghemoglobins

Cited by 72 publications

References 45 publications

Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules

Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules

Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA.

Large-Scale Analysis of Putative Soybean Regulatory Gene Expression Identifies a Myb Gene Involved in Soybean Nodule Development

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