title: APO1 promotes the splicing of chloroplast group II introns and harbors a plant-specific zinc-dependent RNA binding domain.
abstract: Arabidopsis thaliana APO1 is required for the accumulation of the chloroplast photosystem I and NADH dehydrogenase complexes and had been proposed to facilitate the incorporation of [4Fe-4S] clusters into these complexes. The identification of maize (Zea mays) APO1 in coimmunoprecipitates with a protein involved in chloroplast RNA splicing prompted us to investigate a role for APO1 in splicing. We show here that APO1 promotes the splicing of several chloroplast group II introns: in Arabidopsis apo1 mutants, ycf3-intron 2 remains completely unspliced, petD intron splicing is strongly reduced, and the splicing of several other introns is compromised. These splicing defects can account for the loss of photosynthetic complexes in apo1 mutants. Recombinant APO1 from both maize and Arabidopsis binds RNA with high affinity in vitro, demonstrating that DUF794, the domain of unknown function that makes up almost the entirety of APO1, is an RNA binding domain. We provide evidence that DUF794 harbors two motifs that resemble zinc fingers, that these bind zinc, and that they are essential for APO1 function. DUF794 is found in a plant-specific protein family whose members are all predicted to localize to mitochondria or chloroplasts. Thus, DUF794 adds a new example to the repertoire of plant-specific RNA binding domains that emerged as a product of nuclear-organellar coevolution.
authors.affiliation: Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: Watkins
authors.forename: Margarita
authors.initials: M
authors.lastname: Rojas
authors.forename: Giulia
authors.initials: G
authors.lastname: Friso
authors.forename: Klaas J
authors.initials: KJ
authors.lastname: van Wijk
authors.forename: Jörg
authors.initials: J
authors.lastname: Meurer
authors.forename: Alice
authors.initials: A
authors.lastname: Barkan
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D007438:Introns / Q000235:genetics*; D010802:Phylogeny; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003313:Zea mays / Q000235:genetics
chemical_list: C497731:APO1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C017800:zinc-binding protein