Expression and purification of Rhizobium leguminosarum NodD

Feng, Jie; Li, Feng Qing; Li, Qiang; Hu, Hai Liang; Ge, Hong

doi:10.1016/s1046-5928(02)00535-1

Cited by 9 publications

(5 citation statements)

References 14 publications

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“…The NodD proteins of several species are said to be membrane associated, adding further challenges in purification (443). Recently, however, NodD of R. leguminosarum was purified to homogeneity in a soluble form, which may lead to further biochemical and structural studies (134).…”

Section: Host Detection During Nodule Formationmentioning

confidence: 99%

Detection of and Response to Signals Involved in Host-Microbe Interactions by Plant-Associated Bacteria

Brencic

Winans

2005

Microbiol Mol Biol Rev

339

208

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Diverse interactions between hosts and microbes are initiated by the detection of host-released chemical signals. Detection of these signals leads to altered patterns of gene expression that culminate in specific and adaptive changes in bacterial physiology that are required for these associations. This concept was first demonstrated for the members of the family Rhizobiaceae and was later found to apply to many other plant-associated bacteria as well as to microbes that colonize human and animal hosts. The family Rhizobiaceae includes various genera of rhizobia as well as species of Agrobacterium. Rhizobia are symbionts of legumes, which fix nitrogen within root nodules, while Agrobacterium tumefaciens is a pathogen that causes crown gall tumors on a wide variety of plants. The plant-released signals that are recognized by these bacteria are low-molecular-weight, diffusible molecules and are detected by the bacteria through specific receptor proteins. Similar phenomena are observed with other plant pathogens, including Pseudomonas syringae, Ralstonia solanacearum, and Erwinia spp., although here the signals and signal receptors are not as well defined. In some cases, nutritional conditions such as iron limitation or the lack of nitrogen sources seem to provide a significant cue. While much has been learned about the process of host detection over the past 20 years, our knowledge is far from being complete. The complex nature of the plant-microbe interactions makes it extremely challenging to gain a comprehensive picture of host detection in natural environments, and thus many signals and signal recognition systems remain to be described

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Section: Host Detection During Nodule Formationmentioning

confidence: 99%

Detection of and Response to Signals Involved in Host-Microbe Interactions by Plant-Associated Bacteria

Brencic

Winans

2005

Microbiol Mol Biol Rev

339

208

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“…Overexpression of NolR protein in 8401R pU 3 R was constructed by inserting the NolR gene into plasmid pKNDT to replace the nodD gene [24]. R1E and R2E primers were used to amplify the NolR gene.…”

Section: Methodsmentioning

confidence: 99%

Symbiotic plasmid is required for NolR to fully repress nodulation genes inRhizobium leguminosarumA34

Hou

Hong

2008

Acta Biochimica Et Biophysica Sinica

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NolR is a regulator of nodulation genes present in Rhizobium and Sinorhizobium. However, the mechanism by which NolR participates in the inducible transcription of nodulation genes remains unclear. To investigate whether there are other factors regulating the function of NolR, an insertion mutant of NolR in Rhizobium leguminosarum strain 8401, which lacks the symbiotic plasmid, was constructed by homologous recombination. We investigated the effects of NolR inactivation on the expression of nodulation genes. Three inducible nodulation genes (nodA, nodF and nodM) were expressed constitutively in NolR- mutant, MR114. Our results suggested that the symbiotic plasmid is required for NolR to fully repress nodulation genes in Rhizobium leguminosarum A34. In addition, MR114 has provided a useful tool for further study of molecular interactions between NolR and other factors.

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“…The lysate was centrifuged at 19,500 g for 10 min to remove unbroken cells and cell debris. The resulting supernatants were precipitated by 20 -40% saturated ammonium sulfate [36]. After dialysis, the volume of the wild type, I1, I2, D1, and D2 was adjusted to 10 ml; the volume of I3, I4, I5, and I6 was 2 ml and the volume of D3, D4, D5, and D6 was 0.5 ml.…”

Section: Dna Bending By Circular Permutation Assaymentioning

confidence: 99%

The properties of NodD were affected by mere variation in length within its hinge region

Hou¹,

Li²,

Yang³

et al. 2009

ABBS

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In Rhizobium leguminosarum bv. viciae, NodD, a member of the LysR-type transcriptional regulators, while auto-regulating, activates transcription of other nod genes in the presence of naringenin. A hinge region of NodD was previously identified in our laboratory as a functional region independent of its N-terminal DNA-binding and C-terminal regulatory domain. Further study was carried out to see the possible effect of the length variation in the hinge region on NodD's properties. To our surprise, as many as seven classes of phenotypes were observed. Class I is deficient of activating nodA transcription and abolishes auto-regulation; class II is able to activate nodA transcription independently of naringenin and abolishes auto-regulation; class III retains autoregulating but partial activating ability; class IV is able to activate transcription independently of naringenin and retains auto-regulation; in class V, nodA is transcribed constitutively but the transcription level is drastically down-regulated in the presence of naringenin; in class VI, nodA is transcribed constitutively with higher induction ratio; in class VII, nodA is transcribed constitutively with lower induction ratio. To learn more about the possible mechanism, circular permutation assays were done, which showed that the length variation of the hinge of NodD caused by mutation led to the change in bend angles of nod promoter. This finding should help to get an insight into how transcriptional regulation is mediated by NodD at the molecular level as well as to understand the regulatory system of this important family.

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Expression and purification of Rhizobium leguminosarum NodD

Cited by 9 publications

References 14 publications

Detection of and Response to Signals Involved in Host-Microbe Interactions by Plant-Associated Bacteria

Detection of and Response to Signals Involved in Host-Microbe Interactions by Plant-Associated Bacteria

Symbiotic plasmid is required for NolR to fully repress nodulation genes inRhizobium leguminosarumA34

The properties of NodD were affected by mere variation in length within its hinge region

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