Journal Club 1 Questions Bio102: To be handed in at the beginning of class

Loisel et al, 1999, Science.

Name:

1.What are the minimal components required to induce the movement of the
intracellular parasite, Listeria monocytogenes, within cell free extracts.
The minimal components for actin-based movement are the rapid turnover of actin
filaments and require Arp2/3, ADF, and Capping Protein. However, this is the bare
minimum and lead very small movement.

2.What is the maximum velocity of L. monocytogenes in cell free extracts and what
powers this movement?
The maximum velocity 3.3 micrometers per minute. It is powered by using the free
energy released by atp hydrolysis to actin assembly.

3.The predominant forms of actin inside a cell are ATP-G-actin and ADP-F-actin,
Explain how the interconversion of the nucleotide state is coupled to the assembly
and disassembly of actin subunits.

G-actin binds with ATP which then is added to a filaments barbed end of F-actin. F-
actin ADP is simultaneously disassembled where ADP changes to ATP, creating the
treadmilling effect.

4.What would be the consequence for actin filament assembly/disassembly if a

mutation prevented
actins ability to bind ATP?

If there was a mutation such that G-actin could not bind to atp then the filament
could still be formed as atp is not required for formation. It can be formed with ADP
to instigate polymerization. (according to wiki
https://en.wikipedia.org/wiki/Actin#Assembly_dynamics) But it is to believed that
the polymer chain could not be formed as G-actin requires atp to bind to the
filaments barbed end.
5.What would be the consequence if a mutation prevented actins ability to
hydrolyze ATP?
The Listeria would not be motile as it uses the released energy from hydrolyzed atp,
and the end of the polymer chain cannot be disassembled.