Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation

Abstract

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Schizosaccharomyces pombe as a fundamental model for research on mitochondrial gene expression: Progress, achievements and outlooks

“…100 Tandem affinity purification (TAP) of Ppr10 recovered every mt-mRNAs, several mt-LSU and mt-SSU proteins and a novel nuclear-encoded, mitochondrial-localised protein Mpa1. 65,101 Consistently, both Ppr10 and Mpa1 cosedimented with the assembled mitoribosome. Deletion of the two central PPR motifs in Ppr10 compromised its interaction with Mpa1.…”

“…In addition, Ppr10 co‐immunoprecipitated with the mitochondrial translation initiation factor Mti2 (discussed above). Upon either ppr10 or mpa1 deletion, Mti2 (and Mti3) no longer co‐sedimented with the mt‐SSU 101 . Taken together, these data suggested that Ppr10 and Mpa1 cooperatively enhance mitochondrial translation initiation, by physically connecting Mti2 and Mti3 to the mt‐SSU.…”

“…Moreover, mature cytb and cox1 mRNA were less abundant, a result of deprived intron‐encoded maturases 100 . Tandem affinity purification (TAP) of Ppr10 recovered every mt‐mRNAs, several mt‐LSU and mt‐SSU proteins and a novel nuclear‐encoded, mitochondrial‐localised protein Mpa1 65,101 . Consistently, both Ppr10 and Mpa1 co‐sedimented with the assembled mitoribosome.…”

Schizosaccharomyces pombe Sls1 is primarily required for cox1 mRNA translation

“…Mitochondrial proteins were separated using SDS‐PAGE and then electrophoretically transferred to nitrocellulose membranes. The membrane was probed with rabbit polyclonal antibodies (Abs), which were produced as described (Luo et al, 2021; Wang et al, 2017). The secondary Ab used was IRDye 800CW‐conjugated goat anti‐rabbit Abs (LI‐COR Biosciences).…”

“…In S. pombe , PPR protein Ppr2 is required for overall mitochondrial protein synthesis (Kuhl et al, 2011). Ppr10 forms a complex with the translation activator Mpa1 and plays a role in the initiation of mitochondrial protein translation by promoting the binding of the mitochondrial translation initiation factors Mti2 and Mti3 to the small subunit of the mitoribosome (Luo et al, 2021; Wang et al, 2017). In humans, leucine‐rich PPR‐containing protein (LRPPRC) is required for the polyadenylation and translation of mitochondrial mRNAs (mt‐mRNAs) with its partner SRA stem‐loop interacting RNA‐binding protein (SLIRP) (Chujo et al, 2012; Ruzzenente et al, 2012; Sasarman et al, 2010).…”