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The DEAD-box RNA helicase ZmRH48 is required for the splicing of multiple mitochondrial introns,mitochondrial complex biosynthesis,and seed development in maize

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Journal of Integrative Plant Biology 2023年第11期65卷 2456-2468页

作者： Yan-Zhuo Yang Shuo Ding Xin-Yuan Liu Chunhui Xu Feng Sun Bao-Cai Tan Key Laboratory of Plant Development and Environmental Adaptation Biology Ministry of EducationSchool of Life SciencesShandong UniversityQingdao 266237China

RNA helicases participate in nearly all aspects of RNA metabolism by rearranging RNAs or RNA–protein complexes in an adenosine triphosphatedependent manner.Due to the large RNA helicase families in plants,the precise roles of many RNA helicases in plant physiology and development remain to be clarified.Here,we show that mutations in maize(Zea mays)DEAD-box RNA helicase48(Zm RH48)impair the splicing of mitochondrial introns,mitochondrial complex biosynthesis,and seed development.Loss of Zm RH48 function severely arrested embryogenesis and endosperm development,leading to defective kernel formation.Zm RH48 is targeted to mitochondria,where its deficiency dramatically reduced the splicing efficiency of five cis-introns(nad5 intron 1;nad7 introns 1,2,and 3;and ccm Fc intron 1)and one trans-intron(nad2 intron 2),leading to lower levels of mitochondrial complexes I andⅢ.Zm RH48 interacts with two unique pentatricopeptide repeat(PPR)proteins,PPR-SMR1 and SPR2,which are required for the splicing of over half of all mitochondrial introns.PPR-SMR1 interacts with SPR2,and both proteins interact with P-type PPR proteins and Zm-m CSF1 to facilitate intron splicing.These results suggest that Zm RH48 is likely a component of a splicing complex and is critical for mitochondrial complex biosynthesis and seed development.

关键词： intron splicing maize mitochondria RNA helicase seed development ZmRH48

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The Pentatricopeptide Repeat Protein SOT5/EMB2279 is Required for Plastid rpl2 and trnK intron splicing

The Pentatricopeptide Repeat Protein SOT5/EMB2279 is Require...

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第七届长三角植物科学研讨会暨青年学术报告会

作者： Weihua Huang Yajuan Zhu Wenjuan Wu Xuan Li Delin Zhang Ping Yin Jirong Huang Department of Biology College of Life and Environmental SciencesShanghai Normal University Institute of Plant Physiology and Ecology Shanghai Institutes for Biological SciencesChinese Academy of Sciences National Key Laboratory of Crop Genetic Improvement National Centre of Plant Gene ResearchHuazhong Agricultural University

Chloroplast biogenesis and development are highly complex processes requiring interaction between plastid and nuclear genomic products.Using a high-throughput screen for chloroplast biogenesis suppressors in Arabidopsis thaliana,we identified a suppressor of thf1(sot5) that displays virescent and serrated leaves.Further characterization revealed that sot5 mutants are defective in leaf adaxial and abaxial polarity,and act as enhancer of asymmetric leaves2.Map-based cloning identified SOT5 as a gene previously named EMB2279 that encodes a plastid-targeted pentatricopeptide repeat(PPR) protein with 11 PPR motifs.A G-to-A mutation in sot5 leads to a significant decrease in splicing efficiency,generating two additional mRNA variants.As previously reported,the sot5 null mutation is embryo lethal.SOT5 is predicted to bind to specific RNA sequences found in plastid rpl2 and trnK genes,and we found decreased splicing efficiency of the rpl2 and trnK genes in sot5 mutants.Together,our results reveal that the PPR protein SOT5/EMB2279 is required for intron splicing of plastid rpl2 and trnK,providing insights into the role of plastid translation in the coupled development between chloroplasts and leaves.

关键词： Arabidopsis PPR protein plastid gene intron splicing

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PPR17, PPR20 and CRS1 mediate intron splicing by forming a complex in maize chloroplasts

PPR17, PPR20 and CRS1 mediate intron splicing by forming a c...

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2016年全国植物生物学大会

作者： Rui Liu Xiaoyan Zhang Shikai Cao Bao-Cai Tan Key Lab of Plant Cell Engineering and Germplasm Innovation Ministry of EducationSchool of Life SciencesShandong University

In mitochondria and chloroplasts of higher plants,the loss of self-splicing activity of degenerate group II introns results in recruitment of intron splicing factors encoded in the nucleus. One family of such factors is the pentatricopeptide repeat proteins(PPR) which are implicated in editing,splicing,end maturation and regulation of translation in organellar transcripts. PPR proteins belong to a large family with more than 500 members in flowering plants. Because of its large size and frequent embryo lethality in KO mutants,functions of many PPR proteins remain unknown. Here we report the function identification of PPR17 and PPR20 and their physical interactions with Chloroplast RNA Spl icing 1(CRS1) in maize. Null mutation of PPR17 and PPR20 showed similar arrest in embryogenesis at the transition stage. Both PPR17 and PPR20 are P-type PPR proteins with 11 and 17 PPR motifs respectively. Subcellular localization revealed that both proteins are localized in the chloroplast. PPR17 and PPR20 are expressed in all the tissues examined. Analysis of chloroplast transcripts revealed that loss of either the PPR17 or the PPR20 function abolishes the splicing of the atp F and rpl2 introns,suggesting that PPR17 and PPR20 are required for atp F and rpl2 intron splicing. Previous studies have identified that CRM protein CRS1 is also required for atp F intron splicing(Jenkins et al.,1997;Till et al.,2001). Yeast two hybridization,Pull-down and Bi FC analyses indicated that PPR17,PPR20 and CRS1 interact physically. Domain deletion analysis showed that the first four PPR motifs of PPR17 interact with the first two CRM domains of CRS1 and PPR20,and the C-terminal region of CRS1 interacts with PPR20. These results demonstrate that PPR17,PPR20 and CRS1 mediate the atp F intron splicing by forming a complex in maize chloroplasts.

关键词： PPR intron splicing chloroplast embryo and endosperm maize

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The pentatricopeptide repeat protein EMB1270 interacts with CFM2 to splice specific group Ⅱ introns in Arabidopsis chloroplasts

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Journal of Integrative Plant Biology 2021年第11期63卷 1952-1966页

作者： Li Zhang Jingli Chen Liqun Zhang Ying Wei Yajuan Li Xinyun Xu Hui Wu Zhong‐Nan Yang Jirong Huang Fenhong Hu Weihua Huang Yong‐Lan Cui Shanghai Key Laboratory of Plant Molecular Sciences Development Center of Plant Germplasm ResourcesCollege of Life SciencesShanghai Normal UniversityShanghai 200234China

Chloroplast biogenesis requires the coordinated expression of chloroplast and nuclear genes.Here, we show that EMB1270, a plastid-localized pentatricopeptide repeat(PPR) protein, is required for chloroplast biogenesis in Arabidopsis thaliana.Knockout of EMB1270 led to embryo arrest,whereas a mild knockdown mutant of EMB1270 displayed a virescent phenotype. Almost no photosynthetic proteins accumulated in the albino emb1270 knockout mutant. By contrast, in the emb1270 knockdown mutant, the levels of ClpP1 and photosystem I(PSI) subunits were significantly reduced, whereas the levels of photosystem II(PSII) subunits were normal. Furthermore, the splicing efficiencies of the clpP1.2,ycf3.1, ndhA, and ndhB plastid introns were dramatically reduced in both emb1270 mutants. RNA immunoprecipitation revealed that EMB1270 associated with these introns in vivo. In an RNA electrophoretic mobility shift assay(REMSA), a truncated EMB1270 protein containing the 11 Nterminal PPR motifs bound to the predicted sequences of the clpP1.2, ycf3.1, and ndhA introns.In addition, EMB1270 specifically interacted with CRM Family Member 2(CFM2). Given that CFM2 is known to be required for splicing the same plastid RNAs, our results suggest that EMB1270 associates with CFM2 to facilitate the splicing of specific group II introns in Arabidopsis.

关键词： Arabidopsis thaliana CRM Family Member 2 chloroplast EMB1270 intron splicing pentatricopeptide repeat protein

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