Lecture 1 - Biogenesis

Why do we have organelle?

- Compartmentalisation
- Different reactions and their specificity

Prokaryotic cells:

- Don’t have organelles

- Double membrane and cell wall; between them we have a periplasmic space (bacteria is secreting it’s
proteins into)
- No nucleus
- Has a nucleoid
- One chromosome and multiple plasmids

What happened between prokaryotes and eukaryotes to make them (eukaryotes) more complex?

- Evolution
- Need to perform more complex functions
- Larger proteins, different organisation, different function, different arrangements (eg. Different pH)
- Compartmentalisation makes the organisation of eukaryotic cells so much more complex than
prokaryotic: Different processes require different environments, e.g. the low pH of a lysosome (H+
ATPase pumps). Eukaryotic cells need the additional membrane surface area inside the cell e.g. in
mitochondria synthesis of ATP by the f type ATPase (inner mitochondrial membrane)
- The regulation of these processes also needs to be very efficient due to all that is happening in the cell.
- Translation of integral membrane proteins, which are very important in regulating what enters and exits
the cell, must be inserted into a membrane whilst they are translated - this happens at the endoplasmic
reticulum. However, the disadvantage of all this is that there needs to be a complex system of ensuring
that the right vesicles get to the right membranes etc. and this involves a lot of signals. (TIGHT
REGULATION)

How are the compartments organised in respect to one another and how the signalling pathways occur?
(Eukaryotic cell structure)

Nucleus:

- Bound by a double membrane

- Inside the nucleus you have regions of nucleoli (where the ribosomal RNA is made)
- The nuclear DNA is organised into compact chromatin
- Nuclear pore complex around the nucleus which allow molecules to get in and out of the nucleus (tightly
controlled)
- The nuclear membrane is continuous with the endoplasmic reticulum membrane
- Also has the rough endoplasmic reticulum attached as well (ER with the ribosomes attached)
- chromatin composed of DNA and proteins; site of tRNA and mRNA synthesis
- nucleoli as sites of rRNA synthesis and ribosome assembly (also cytoplasm)

ER:

- Connected with the nuclear membrane

- The proteins when they come out of the nucleus get translated into the ER (efficient)
- Proteins are produced in the ER and some need to leave the ER to get to their destination so the signals
from the ER tell them where to go
 - Some proteins are then budded off in vesicles for transported to different places, they might be
transported outside the cell to fuse with the plasma membrane allowing the content to enter the
bloodstream BUT they need to go through the golgi apparatus

Ribosomes:

- Large subunit and a smaller subunit, RNA and from that a peptide being made.
- Ribosomal distribution throughout the cell cycle: this is performed by placing a dye that intercalates with
the DNA and measuring how much it fluoresces.

How is the endoplasmic reticulum inherited?

In the interphases it is distributed through out the cytoplasm and its structure is organised by a
microtubule’s cytoskeleton. When the mitosis phase starts at the prophase part; phosphorylation event
of the nucleus scaffolding proteins; which causes the nuclear membrane to breakdown and the
microtubules that support the ER start to tear. Basically, gets broken down into little fragments during
metaphase. This leads to an equal distribution of the ER fragments until finally its structure is established
in the G1 phase

Golgi Apparatus:

- The proteins from the ER are sent here for further modifications
- Then these cells can be secreted outside the cell

How is the Golgi apparatus inherited?

The Golgi forms a ribbon in the interphase; then at the start of prophase it begins to fragment. In meta phase
it has fragmented into so many fragments (50 to 70nm vesicles). Then in anaphase these fragments are
equally distribution of these contents In G1, the organisation is maintained; creating the Golgi ribbon again at
the periphery of the nucleus.
Lysosomes:

- After the ER or the golgi apparatus, some proteins go to the lysosomes.

- The enzymes may become degradative and work in a low pH

Mitochondria:

- Double membrane bound organelle

Peroxisomes:

- Fatty acid breakdown takes place

Plasma cell: (White blood cell that has differentiated)

- Plasma cell function is to make a lot of antibodies

- The antibodies need to be secreted outside the cell, which is why they need to be assembled in the
endoplasmic reticulum and then they need to be secreted outside the cell.

How is the endoplasmic reticulum inherited?

In the interphases it is distributed through out the cytoplasm and its structure is organised by a microtubule’s
cytoskeleton. When the mitosis phase starts at the prophase part; phosphorylation event of the nucleus
scaffolding proteins; which causes the nuclear membrane to breakdown and the microtubules that support the ER
start to tear. Basically, gets broken down into little fragments during metaphase. This leads to an equal
distribution of the ER fragments until finally its structure is established in the G1 phase
Learning Objectives:

1. describe organisation, function and inheritance of eukaryotic organelles describe how the regulation of
the cell cycle is crucial for cellular division
2. describe the mechanism of protein targeting and sorting into organelles
3. discuss the complexity of cellular compartmentalisation and the co-ordination of signalling pathways
4. describe experimental approaches that have been used to gain knowledge in organelle biogenesis and the
organisation of the cell cycle
Learning objectives

1. Define and describe conjugation and the steps involved

2. Understand and describe the process of plasmid mobilisation