Dataset Information

Mass Spectrometry of Escherichia coli expressing ParA from F' plasmid or weak IPTG inducible promoter (pDSW210 vector)

ABSTRACT: Comparisonof ParA levels in E. coli carrying mini-F plasmids (ParA expressed from the native ParA promoter) with ParA-GFP levels expressed froma weak Ptrc promoter in pDSW210 vector.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Escherichia Coli

SUBMITTER: RAMANUJAM SRINIVASAN

LAB HEAD: RAMANUJAM SRINIVASAN

PROVIDER: PXD064422 | Pride | 2025-07-07

REPOSITORIES: Pride

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Publications

Mutational analysis of the F plasmid partitioning protein ParA reveals residues required for oligomerization and plasmid maintenance.

Mitra Nivedita N Mishra Dipika D Mudaliyar Manasi M Yadav Rinku R Zinjurte Suyog S Puthethu Irene Aniyan IA Gayathri Pananghat P Ghosal Debnath D Srinivasan Ramanujam R

Nucleic acids research 20250601 12

Mobile genetic elements such as plasmids play a crucial role in the emergence of antimicrobial resistance. Hence, plasmid maintenance proteins such as ParA of the Walker A-type ATPases/ParA superfamily are potential targets for novel antibiotics. Plasmid partitioning by ParA relies on ATP-dependent dimerization and formation of chemophoretic gradients of ParA-ATP on bacterial nucleoids. Though polymerization of ParA has been reported in many instances, the need for polymerization in plasmid main ...[more]

PMID: 40568943

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Project description:G protein coupled receptor (GPCR) signaling in osteoblasts (OBs) is an important regulator of bone formation. We previously described a mouse model expressing Rs1, an engineered constitutively active Gs-coupled GPCR, under the control of the 2.3 kb-Col I promoter. These mice showed a dramatic age-dependent increase in trabecular bone which were accompanied by an increase in OB lineage cells, especially immature OBs, indicated by an expansion of cells expressing Osterix and Runx2 in the whole femur. In this study, we further evaluated how Gs signaling in OBs affects intramembranous bone formation by examining calvariae of one-and nine-week-old Col1(2.3)/Rs1 mice. Rs1 calvariae displayed a dramatic increase in total volume and trabecular bone volume with partial loss of cortical structure. By immunohistochemistry, Osterix was detected in cells throughout the inter-trabecular space in Rs1 expressing mice while Osteocalcin was expressed predominantly in cells along bone surfaces. These findings resembled that previously seen in Rs1 femoral bones, suggesting the role of paracrine mediators secreted from OBs driven by 2.3 kb-Col I promoter could influence early OB commitment, differentiation, and/or proliferation. However, it is still unclear how G protein signaling in mature OBs leads to the observed alterations in bone mass. In this study, we investigated the cellular basis of the skeletal changes by assessing the effect of Rs1 expression in vivo on the transcriptome of mature OBs. We identified the complete set of Gs-GPCRs and other GPCRs that are expressed on OBs which may contribute to the observed skeletal phenotype. Candidate paracrine mediators of the effect of Gs signaling in OBs were determined. Genes affected by Rs1 signaling include those encoding proteins important for cell differentiation, cytokines and growth factors, angiogenesis, coagulation, and energy metabolism. Our results identify novel candidate mediators of the anabolic response to the skeleton to Gs signaling in mature OBs. We utilized a microarray approach to examine Rs1-induced alterations in the OB transcriptome. The approach used to identify an approximate snapshot of the OB transcriptome at the time of sacrifice by isolating the OB population that expresses Rs1 by GFP labeling, without the use of cell culture. We compared gene expression between control OBs and OBs-expressing Rs1 in triplicates.

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