N‐terminal targeting sequences and coding sequences act in concert to determine the localization and trafficking pathway of apicoplast proteins in Toxoplasma gondii.

Backgound Information: Toxoplasma gondii has a relict plastid, the apicoplast, to which nuclear‐encoded proteins are targeted after synthesis in the cytosol. Proteins exclusively found in the apicoplast use a Golgi‐independent route for trafficking, while dually targeted proteins found in both the apicoplast and the mitochondrion use a Golgi‐dependent route. For apicoplast targeting, N‐terminal signal sequences have been shown to direct the localization of different reporters. In this study, we use chimeric proteins to dissect out the roles of N‐terminal sequences and coding sequences in apicoplast localization and the choice of the trafficking route. Results: We show that when the N‐termini of a dually targeted protein, TgTPx1/2, or of an apicoplast protein, TgACP, are fused with the reporter protein, enhanced green fluorescent protein (eGFP) or endogenous proteins, TgSOD2, TgSOD3, TgACP, or TgTPx1/2, the chimeric proteins exhibit flexibility in apicoplast targeting depending on the coding sequences. Further, the chimeras that are localized to the apicoplast use different trafficking pathways depending on the combination of the N‐terminal signals and the coding sequences. Conclusion and Significance: This report shows, for the first time, that in addition to the N‐terminal signal sequences, targeting and trafficking signals also reside within the coding sequences of apicoplast proteins. [ABSTRACT FROM AUTHOR]