The art of multi-tasking: replicating DNA in a multi-nucleated malaria parasite
Supervisors:
Dr Abhinay Ramaprasad, University of Glasgow
Prof Mick Urbaniak, Lancaster University
Prof Richard McCulloch, University of Glasgow
Project Summary:
Malaria parasites proliferate and cause disease within red blood cells through a remarkable and unique process of cell division known as schizogony. Unlike most eukaryotes, the parasite first undergoes multiple rounds of DNA replication and nuclear division without cytokinesis, producing a multi-nucleated form called the schizont. Uniquely, DNA in individual nuclei within the same parasite replicate only once but independently at markedly different times.
Mechanisms that contribute to this asynchrony are completely unknown. In model eukaryotes, a pre-replication complex (pre-RC) assembles at origins of DNA replication, which then gets activated when it is time to start DNA replication. Whilst some core proteins of this complex are conserved in Plasmodium falciparum, the deadliest of human malaria species, other factors have diverged significantly or are considered absent altogether.
What is the composition of a malarial pre-RC? Which of them are crucial for faithful DNA replication? And what are their dynamics in a multinucleated Plasmodium schizont? These are the key questions this PhD project seeks to address.
You will use CRISPR-Cas9 and DiCre technologies to create inducible knockouts of pre-RC proteins. You will use advanced imaging and time-lapse microscopy to study and characterise the defects in the resulting mutant parasites. To dissect the underlying molecular mechanisms, you will also use -omics approaches such as co-immunoprecipitation coupled mass spectrometry and Nanopore sequencing.
If you are intrigued by the challenge of exploring
fundamental questions in an unconventional parasitic eukaryote, we encourage you to apply.