The Hippo kinase cascade regulates a contractile cell behavior and cell density in a close unicellular relative of animals.
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The coHpo gene targeted for knockout is CAOG_01932.
"target the coHpo gene (CAOG_01932)"
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Only one of three predicted coHpo isoforms encodes the SARAH domain, and that isoform was used for the rescue transgene.
"There are three predicted isoforms of the coHpo sequence, but only one of these encodes the SARAH domain, which is important for Hippo kinase activity in Hippo pathway signaling"
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Loss of coHpo increases nuclear localization of coYki.
"In contrast, the majority of coHpo-/- and coWts-/- cells showed nuclear levels of mScarlet-coYki greater than or equal to that seen in the cytoplasm"
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coHpo loss does not change proliferation in adherent culture.
"In adherent growth conditions, coWts-/- and coHpo-/- cells proliferated at similar rates as WT cells"
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The elongated (contractile) cell phenotype is rescued by a coHpo transgene.
"The presence of elongated cells was rescued by transgenic expression of coHpo or coWts in the respective mutant"
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coHpo loss increases cell packing density within aggregates.
"These results indicate that loss of the Hippo pathway kinases increases cell density within Capsaspora aggregates."
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coHpo-/- aggregates have normal size and circularity.
"coHpo -/- aggregates showed similar size and circularity as WT aggregates"
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A non-phosphorylatable coYki phenocopies the kinase mutants; double-mutant epistasis was not possible.
"Since techniques are currently unavailable to create double mutants in Capsaspora, we could not test this possibility through classic genetic epistasis."