Cellular localization of choline-utilization proteins in Streptococcus pneumoniae using novel fluorescent reporter systems

Abstract:

The molecular mechanisms underlying cell growth, cell division and pathogenesis in Streptococcus pneumoniae are still not fully understood. Single-cell methodologies are potentially of great value to investigate S. pneumoniae cell biology. Here, we report the construction of novel plasmids for single and double cross-over integration of functional fusions to the gene encoding a fast folding variant of the green fluorescent protein (GFP) into the S. pneumoniae chromosome. We have also established a zinc-inducible system for the fine control of gfp-fusion gene expression and for protein depletion experiments in S. pneumoniae. Using this novel single cell toolkit, we have examined the cellular localization of the proteins involved in the essential process of choline decoration of S. pneumoniae teichoic acid. GFP fusions to LicA and LicC, enzymes involved in the activation of choline, showed a cytoplasmic distribution, as predicted from their primary sequences. A GFP fusion to the choline importer protein LicB showed clear membrane localization. GFP fusions to LicD1 and LicD2, enzymes responsible for loading of teichoic acid subunits with choline, are also membrane-associated, even though both proteins lack any obvious membrane spanning domain. These results indicate that the decoration of teichoic acid by the LicD enzymes is a membrane-associated process presumably occurring at lipid-linked teichoic acid precursors.

SEEK ID: https://fairdomhub.org/publications/106

PubMed ID: 19737355

Projects: Noisy-Strep

Publication type: Not specified

Journal: Mol. Microbiol.

Citation:

Date Published: 8th Sep 2009

Registered Mode: Not specified

Authors: Alice Eberhardt, Ling J Wu, Jeff Errington, Waldemar Vollmer,

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Created: 10th Feb 2011 at 10:27

Last updated: 8th Dec 2022 at 17:26

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