Studies of cytoskeletal proteins FilP, Scy and CtpS in Streptomyces venezuelae
(2014) MOBT18 20132Degree Projects in Molecular Biology
- Abstract
- Abstract
The cytoskeleton allows for complex organization of cellular components and one important manifestation of this is the ability for cells to have polar growth. The recently discovered bacterial cytoskeleton has led to new lines of investigation, where information from bacterial model systems can not only lead to interesting and important discoveries in bacterial cells, but also further the understanding of the eukaryotic cytoskeleton. One bacterial model organism that has been important in the elucidation of the function of several cytoskeletal proteins, including FilP and Scy, is Streptomyces coelicolor. A new model organism in studies of cytoskeletal proteins is Streptomyces venezuelae, which has many properties making it a... (More) - Abstract
The cytoskeleton allows for complex organization of cellular components and one important manifestation of this is the ability for cells to have polar growth. The recently discovered bacterial cytoskeleton has led to new lines of investigation, where information from bacterial model systems can not only lead to interesting and important discoveries in bacterial cells, but also further the understanding of the eukaryotic cytoskeleton. One bacterial model organism that has been important in the elucidation of the function of several cytoskeletal proteins, including FilP and Scy, is Streptomyces coelicolor. A new model organism in studies of cytoskeletal proteins is Streptomyces venezuelae, which has many properties making it a viable candidate for further understanding of the functions of these proteins. Further investigation into the interaction partners of these known cytoskeletal proteins is essential for understanding the molecular mechanics of the cytoskeleton and polar growth. One suspected interaction partner of the S. venezuelae FilP protein is CTP synthase, CtpS. This thesis aimed to further the understanding of the FilP, Scy and CtpS proteins and their functions. Knock-out mutations of filP and scy in S. venezuelae were investigated, mainly by light microscopy. Further work was done in establishing new genetic tools for use in S. venezuelae. Attempts to clone fluorescently tagged CtpS into S. venezuelae were also done. This is a part of the work required to fully elucidate the functions of these proteins and their importance in the cytoskeleton and polar growth, in particular. (Less) - Abstract
- Popular science summary:
Understanding the bacterial cytoskeleton
The bacterial cytoskeleton was discovered not too long ago. Since its discovery, it has been a source of information and understanding of how this essential cellular system is established and maintained. The cytoskeleton allows for subcellular organization, which is the key behind many cellular functions. One such function is polar growth, which is found in the bacterial Streptomyces species.
The polar growth shown by Streptomyces is very similar to that seen in filamentous fungi, where the bacteria form hyphae, which branch and lead to the formation of a mycelium. How is this form of growth established and maintained? One important finding was the DivIVA protein,... (More) - Popular science summary:
Understanding the bacterial cytoskeleton
The bacterial cytoskeleton was discovered not too long ago. Since its discovery, it has been a source of information and understanding of how this essential cellular system is established and maintained. The cytoskeleton allows for subcellular organization, which is the key behind many cellular functions. One such function is polar growth, which is found in the bacterial Streptomyces species.
The polar growth shown by Streptomyces is very similar to that seen in filamentous fungi, where the bacteria form hyphae, which branch and lead to the formation of a mycelium. How is this form of growth established and maintained? One important finding was the DivIVA protein, which has been shown to establish polar growth after forming protein foci at the emerging hyphal tips.
However, DivIVA does not act alone. Other proteins have been found to aid in polar growth. Two of these proteins, FilP and Scy, have previously been extensively studied in Streptomyces coelicolor. This model organism does, unfortunately, have some properties that make it ill-suited for certain investigations. Therefore, the closely related Streptomyces venezuelae has been brought forth as a new model system, where FilP and Scy can be studied further, to attempt and understand their functions better. To enable use of the new model organism S. venezuelae, novel genetic tools had to be created. To investigate the cytoskeletal functions of FilP and Scy, as well as establish functional genetic tools, in S. venezuelae were the goals of this thesis.
The FilP and Scy proteins are not essential for polar growth in S. venezuelae, but they are important.
FilP and Scy were found to have effects on polar growth in S. venezuelae. The strain lacking the filP gene did, however, not have severe phenotypic effects. Another mutant strain, where the scy gene was knocked out in a similar way showed a more interesting phenotype. The hyphae of this strain were of two distinct types, one healthy and one sick, which has not been seen in the scy knock-out mutant of S. coelicolor. This observed difference between the organisms show the importance of investigations in other model organisms, to further understanding of protein functions.
Finally, a knock-out strain of both filP and scy was constructed, where it was found that the phenotypes had additive effects. This is an indicator that the proteins interact and are important for the functionality of each-other in establishing and maintaining functional polar growth in S. venezuelae. The results presented in this thesis further the knowledge of the functions of the FilP and Scy proteins, as well as establish functional genetic tools for cloning, in S. venezuelae,
Supervisor: Nora Ausmees
Master Degree Project, 60 credits, in Microbiology, 2013-2014
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/4519647
- author
- Fröjd, Markus
- supervisor
-
- Nora Ausmees LU
- organization
- course
- MOBT18 20132
- year
- 2014
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 4519647
- date added to LUP
- 2014-06-26 16:56:51
- date last changed
- 2014-06-26 16:56:51
@misc{4519647,
abstract = {{Popular science summary:
Understanding the bacterial cytoskeleton
The bacterial cytoskeleton was discovered not too long ago. Since its discovery, it has been a source of information and understanding of how this essential cellular system is established and maintained. The cytoskeleton allows for subcellular organization, which is the key behind many cellular functions. One such function is polar growth, which is found in the bacterial Streptomyces species.
The polar growth shown by Streptomyces is very similar to that seen in filamentous fungi, where the bacteria form hyphae, which branch and lead to the formation of a mycelium. How is this form of growth established and maintained? One important finding was the DivIVA protein, which has been shown to establish polar growth after forming protein foci at the emerging hyphal tips.
However, DivIVA does not act alone. Other proteins have been found to aid in polar growth. Two of these proteins, FilP and Scy, have previously been extensively studied in Streptomyces coelicolor. This model organism does, unfortunately, have some properties that make it ill-suited for certain investigations. Therefore, the closely related Streptomyces venezuelae has been brought forth as a new model system, where FilP and Scy can be studied further, to attempt and understand their functions better. To enable use of the new model organism S. venezuelae, novel genetic tools had to be created. To investigate the cytoskeletal functions of FilP and Scy, as well as establish functional genetic tools, in S. venezuelae were the goals of this thesis.
The FilP and Scy proteins are not essential for polar growth in S. venezuelae, but they are important.
FilP and Scy were found to have effects on polar growth in S. venezuelae. The strain lacking the filP gene did, however, not have severe phenotypic effects. Another mutant strain, where the scy gene was knocked out in a similar way showed a more interesting phenotype. The hyphae of this strain were of two distinct types, one healthy and one sick, which has not been seen in the scy knock-out mutant of S. coelicolor. This observed difference between the organisms show the importance of investigations in other model organisms, to further understanding of protein functions.
Finally, a knock-out strain of both filP and scy was constructed, where it was found that the phenotypes had additive effects. This is an indicator that the proteins interact and are important for the functionality of each-other in establishing and maintaining functional polar growth in S. venezuelae. The results presented in this thesis further the knowledge of the functions of the FilP and Scy proteins, as well as establish functional genetic tools for cloning, in S. venezuelae,
Supervisor: Nora Ausmees
Master Degree Project, 60 credits, in Microbiology, 2013-2014
Department of Biology, Lund University}},
author = {{Fröjd, Markus}},
language = {{eng}},
note = {{Student Paper}},
title = {{Studies of cytoskeletal proteins FilP, Scy and CtpS in Streptomyces venezuelae}},
year = {{2014}},
}