Cellular homeostasis of Escherichia coli probed by super-resolution microscopy

Jonas van den Berg

Cellular homeostasis of Escherichia coli probed by super-resolution microscopy

Jonas van den Berg

Enzymologie

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In elke cel zijn er miljoenen macromoleculen, waaronder DNA, RNA en eiwitten, die allemaal in een klein volume moeten verblijven. Stel je voor, dat de cel een keuken is in een groot restaurant. Hier moeten de manager, koks, keukenpersoneel en obers hand in hand werken om de restaurantgasten gauw van eten te voorzien. Daarvoor is het belangrijk om voldoende personeel in dienst te hebben. Met onderbezetting zouden de gasten erg lang moeten wachten op hun voedsel. Omdat de ruimte in elke keuken begrenst is, kunnen maar een beperkt aantal medewerkers worden ingehuurd, zonder dat ze elkaar bij het uitvoeren van hun opdrachten hinderen.
In een cel verlopen processen vergelijkbaar met de werkzaamheden in zo’n keuken. Het aantal moleculen moet worden aangepast aan de celgrootte. Op microscopische schaal zitten de moleculen in een cel erg dicht bij elkaar. Dit fenomeen noemt men macromoleculaire crowding. De mate van crowding kan de effectiviteit van enzymreacties en andere processen beïnvloeden. Het is zeer waarschijnlijk, dat de cel – net zo als de keukenmanager – de eigen ‘volheid’ nauwkeurig regelt.
In dit proefschrift wordt uitgelegd welke mechanismen cellen hebben om de crowding te regelen. Met behulp van fluorescentiemicroscopie is de ‘volheid’ van de cel onder verschillende groei-omstandigheden bepaald. Verder heb ik onderzoek gedaan aan zogenoemde mechanosensitieve kanalen. Dit zijn membraaneiwitten die belangrijk zijn in het reguleren van de concentratie aan osmotisch-actieve stoffen en daarmee het volume van de cel reguleren. Mijn onderzoek ondersteunt in het algemeen de hypothese van een strakke regulering van macromoleculaire crowding.

Originele taal-2	English
Kwalificatie	Doctor of Philosophy
Toekennende instantie	Rijksuniversiteit Groningen
Begeleider(s)/adviseur	Poolman, Berend, Supervisor
Datum van toekenning	28-okt.-2016
Plaats van publicatie	[Groningen]
Uitgever	Rijksuniversiteit Groningen
Gedrukte ISBN's	978-90-367-9179-3
Elektronische ISBN's	978-90-367-9177-9
Status	Published - 2016

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Title and contents Final publisher's version, 287 KB
Chapter 1Final publisher's version, 2,43 MB
Chapter 2Final publisher's version, 2,88 MB
Chapter 3Final publisher's version, 8,13 MB
Chapter 4Final publisher's version, 6,74 MB
Chapter 5Final publisher's version, 1,81 MB
SummaryFinal publisher's version, 88,9 KB
Nederlandse SamenvattingFinal publisher's version, 56,9 KB
Deutsche ZusammenfassungFinal publisher's version, 58 KB
AcknowledgmentsFinal publisher's version, 79,3 KB
Complete thesisFinal publisher's version, 21,8 MB
PropositionsFinal publisher's version, 49,1 KB

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title = "Cellular homeostasis of Escherichia coli probed by super-resolution microscopy",

abstract = "In every cell there are millions of macromolecules, like for example DNA, RNA, and proteins, all of which have to reside in a tiny volume. Imagine the cell as a kitchen in a big restaurant. All the cooks, kitchen helpers and waiters have to work together closely to make sure all the guests are being served in time. The restaurant manager has to hire enough staff to satisfy the customers. On the other hand, if more people are hired than workstations are available, the productivity will go down, because they will only stand in each other{\textquoteright}s way. Inside a cell, the situation is quit similar; the number of macromolecules has to be adjusted to the available cell volume. Microscopically seen, each cell is highly crowded. This phenomenon is called macromolecular crowding and is inherent to any type of cell. Changes in crowding can make certain biochemical processes less efficient, which makes it very likely that the cell – just like the restaurant manager – tries to carefully adjust its {\textquoteleft}fullness{\textquoteright}. In this thesis, possible mechanisms of how macromolecular crowding could be regulated, are described. Fluorescence microscopy was used to measure the {\textquoteleft}fullness{\textquoteright} of a cell under varying growth conditions. In another part, the functionality of so-called {\textquoteleft}mechanosensitive channels{\textquoteright} was studied. These are membrane proteins that regulate the concentration of small molecules in the cytoplasm and thereby the volume of the cell. The experimental proof of this work generally supports the hypothesis of tight regulation of macromolecular crowding.",

author = "{van den Berg}, Jonas",

year = "2016",

language = "English",

isbn = "978-90-367-9179-3",

publisher = "Rijksuniversiteit Groningen",

school = "University of Groningen",

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TY - BOOK

T1 - Cellular homeostasis of Escherichia coli probed by super-resolution microscopy

AU - van den Berg, Jonas

PY - 2016

Y1 - 2016

N2 - In every cell there are millions of macromolecules, like for example DNA, RNA, and proteins, all of which have to reside in a tiny volume. Imagine the cell as a kitchen in a big restaurant. All the cooks, kitchen helpers and waiters have to work together closely to make sure all the guests are being served in time. The restaurant manager has to hire enough staff to satisfy the customers. On the other hand, if more people are hired than workstations are available, the productivity will go down, because they will only stand in each other’s way. Inside a cell, the situation is quit similar; the number of macromolecules has to be adjusted to the available cell volume. Microscopically seen, each cell is highly crowded. This phenomenon is called macromolecular crowding and is inherent to any type of cell. Changes in crowding can make certain biochemical processes less efficient, which makes it very likely that the cell – just like the restaurant manager – tries to carefully adjust its ‘fullness’. In this thesis, possible mechanisms of how macromolecular crowding could be regulated, are described. Fluorescence microscopy was used to measure the ‘fullness’ of a cell under varying growth conditions. In another part, the functionality of so-called ‘mechanosensitive channels’ was studied. These are membrane proteins that regulate the concentration of small molecules in the cytoplasm and thereby the volume of the cell. The experimental proof of this work generally supports the hypothesis of tight regulation of macromolecular crowding.

AB - In every cell there are millions of macromolecules, like for example DNA, RNA, and proteins, all of which have to reside in a tiny volume. Imagine the cell as a kitchen in a big restaurant. All the cooks, kitchen helpers and waiters have to work together closely to make sure all the guests are being served in time. The restaurant manager has to hire enough staff to satisfy the customers. On the other hand, if more people are hired than workstations are available, the productivity will go down, because they will only stand in each other’s way. Inside a cell, the situation is quit similar; the number of macromolecules has to be adjusted to the available cell volume. Microscopically seen, each cell is highly crowded. This phenomenon is called macromolecular crowding and is inherent to any type of cell. Changes in crowding can make certain biochemical processes less efficient, which makes it very likely that the cell – just like the restaurant manager – tries to carefully adjust its ‘fullness’. In this thesis, possible mechanisms of how macromolecular crowding could be regulated, are described. Fluorescence microscopy was used to measure the ‘fullness’ of a cell under varying growth conditions. In another part, the functionality of so-called ‘mechanosensitive channels’ was studied. These are membrane proteins that regulate the concentration of small molecules in the cytoplasm and thereby the volume of the cell. The experimental proof of this work generally supports the hypothesis of tight regulation of macromolecular crowding.

M3 - Thesis fully internal (DIV)

SN - 978-90-367-9179-3

PB - Rijksuniversiteit Groningen

CY - [Groningen]

ER -

Cellular homeostasis of Escherichia coli probed by super-resolution microscopy

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