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Caulobacter crescentus: the genetic mechanisms governing the lipidome, metabolism and cell wall biosynthesis
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Stankeviciute, Gabriele. Caulobacter crescentus: the genetic mechanisms governing the lipidome, metabolism and cell wall biosynthesis. Retrieved from https://doi.org/doi:10.7282/t3-8wpw-7375

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TitleCaulobacter crescentus: the genetic mechanisms governing the lipidome, metabolism and cell wall biosynthesis
NameStankeviciute, Gabriele (author); Klein, Eric A. (chair); Brannigan, Grace (internal member); Shain, Daniel H. (internal member); Huang, Kerwyn Casey (outside member); Rutgers University; Camden Graduate School
Date Created2021
Other Date2021-01 (degree)
SubjectComputational and Integrative Biology
Extent1 online resource (ix, 171 pages)
DescriptionDespite extensive studies that have been done to understand the regulation of ‘prototypical’ cell morphologies, like the classical rod-shaped E. coli, the regulation of the vast majority of unique, non-symmetric cell shapes remains poorly characterized. Certain bacterial species form thin cellular appendages called stalks or ‘prosthecae’ which are at times used for reproduction, adhesion and other purposes. The Alphaproteobacterium Caulobacter crescentus changes its cell body and stalk length in response to phosphate starvation. The following work uses this model organism to illustrate the molecular underpinnings that enable the organism to adapt to stress by changing its cell length and cellular chemical composition via the remodeling of the cell wall, lipid membranes and overall cellular metabolism.

The stalk cell wall was shown to be distinct due to its ability to evade lysozyme degradation and peptidoglycan protein binding. These observations implied that the stalk’s chemical composition and/or the peptidoglycan geometric configuration leads to the lack of digestion and PG recognition. The stalk PG was found to be crosslinked with LD-transpeptidation as opposed to the more common DD-transpeptidation, this was identified to be mediated predominantly by the LD-transpeptidase LtdD. Consistent with the results seen in Caulobacter, lysozyme resistance was found in other species with peptidoglycan enriched with LD-crosslinks.

During phosphate starvation, a considerable increase in lipid demand occurs as lipid membranes expand due to the stalk elongation. We characterized the increase in the production of non-phosphate derived lipids that are made in order to compensate for that increased demand and found that various anionic glycolipids are largely increased to replace the negatively charged phosphatidylglycerol. In the MS analysis we found a novel lipid species, diglycosylated ceramide (HexHexA-Cer) and identified the entire biosynthesis pathway of this unique lipid. We established that in the absence of ceramide, Caulobacter becomes resistant to the antibiotics polymyxin B and more sensitive to killing by bacteriophage Cr30. Using these two fitness phenotypes we were able to develop a two-step screening method to identify genes responsible for ceramide synthesis. Using our genetic screen, we showed this pathway is widespread throughout bacteria and evolved independently of the eukaryotic ceramide synthesis pathway.
 
NotePh.D.
NoteIncludes bibliographical references
Genretheses, ETD doctoral
Persistent URLhttps://doi.org/doi:10.7282/t3-8wpw-7375
LanguageEnglish
CollectionCamden Graduate School Electronic Theses and Dissertations
Organization NameRutgers, The State University of New Jersey
RightsThe author owns the copyright to this work.
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