Abstract
ABSTRACTHow animals emerged from their unicellular ancestor remains a major evolutionary question. New genome data from the closest unicellular relatives of animals have provided important insights into the evolution of animal multicellularity. We know that the unicellular ancestor of animals had an unexpectedly complex genetic repertoire, including many genes that are key to animal development and multicellularity. Thus, assessing the function of these genes among unicellular relatives of animals is key to understanding how they were co-opted at the onset of the Metazoa. However, such analyses have been hampered by the lack of genetic tools. Progress has been made in choanoflagellates and teretosporeans, two of the three lineages closely related to animals, whereas no tools are yet available for functional analysis in the third lineage: the filastereans. Importantly, filastereans have a striking repertoire of genes involved in transcriptional regulation and other developmental processes. Here, we describe a reliable transfection method for the filasterean Capsaspora owczarzaki. We also provide a set of constructs for visualising subcellular structures in live cells. These tools convert Capsaspora into a unique experimentally tractable organism to use to investigate the origin and evolution of animal multicellularity.
Highlights
The transition to animal multicellularity from a single-celled ancestor is one of the most intriguing events in the history of life (King, 2004; Ruiz-Trillo et al, 2007; Rokas, 2008; Knoll, 2011; Richter and King, 2013; Cavalier-Smith, 2017; Sebé-Pedrós et al, 2017)
These include genes involved in cell adhesion, such as those encoding integrins and cadherins, cell-tocell communication, such as those encoding tyrosine kinases, and transcriptional regulation, such as the developmental transcription factor Brachyury (Sebé-Pedrós et al, 2010, 2011; Sebé-Pedrós et al, 2013a; Nichols et al, 2012; Suga et al, 2012)
These findings imply that the co-option of ancestral genes into new functions was an important mechanism for the transition to animal multicellularity
Summary
The transition to animal multicellularity from a single-celled ancestor is one of the most intriguing events in the history of life (King, 2004; Ruiz-Trillo et al, 2007; Rokas, 2008; Knoll, 2011; Richter and King, 2013; Cavalier-Smith, 2017; Sebé-Pedrós et al, 2017). Recent analysis of the genome, transcriptome, proteome and phosphoproteome of the filasterean amoeba Capsaspora owczarzaki (Fig. S1), hereafter Capsaspora, provided important insights into the origins of animal multicellularity and the nature of their unicellular ancestor (Suga et al, 2013; Sebé-Pedrós et al, 2016a,b). RESULTS AND DISCUSSION Capsaspora transfection using calcium phosphate precipitation There are several protocols available for the transient transfection of plasmid DNA for eukaryotic cells.
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