Abstract
The nucleolar protein pescadillo (PES) controls biogenesis of the 60S ribosomal subunit through functional interactions with Block of Proliferation 1 (BOP1) and WD Repeat Domain 12 (WDR12) in plants. In this study, we determined protein characteristics and in planta functions of BOP1 and WDR12, and characterized defects in plant cell growth and proliferation caused by a deficiency of PeBoW (PES-BOP1-WDR12) proteins. Dexamethasone-inducible RNAi of BOP1 and WDR12 caused developmental arrest and premature senescence in Arabidopsis, similar to the phenotype of PES RNAi. Both the N-terminal domain and WD40 repeats of BOP1 and WDR12 were critical for specific associations with 60S/80S ribosomes. In response to nucleolar stress or DNA damage, PeBoW proteins moved from the nucleolus to the nucleoplasm. Kinematic analyses of leaf growth revealed that depletion of PeBoW proteins led to dramatically suppressed cell proliferation, cell expansion, and epidermal pavement cell differentiation. A deficiency in PeBoW proteins resulted in reduced cyclin-dependent kinase Type A activity, causing reduced phosphorylation of histone H1 and retinoblastoma-related (RBR) protein. PeBoW silencing caused rapid transcriptional modulation of cell-cycle genes, including reduction of E2Fa and Cyclin D family genes, and induction of several KRP genes, accompanied by down-regulation of auxin-related genes and up-regulation of jasmonic acid-related genes. Taken together, these results suggest that the PeBoW proteins involved in ribosome biogenesis play a critical role in plant cell growth and survival, and their depletion leads to inhibition of cell-cycle progression, possibly modulated by phytohormone signaling.
Highlights
Gene ID AT5G53300 AT5G14520 AT2G40360 AT5G15550 AT1G07370 AT2G31270 AT3G54710 AT2G29680 AT4G14700 AT4G12620 AT2G37560 AT3G23580 AT2G36010 AT5G22220 AT1G47870 AT3G12280 AT2G23430 AT3G50630 AT5G48820 AT2G32710 AT3G24810 AT3G19150 AT1G49620 AT1G70210 AT1G76540 AT4g34160 AT5G65420 AT4G03270 AT1G07820 AT3G54180 AT1G76540 AT2G44810 AT3G45140 AT5G42650 AT2G06050 AT1G19640 AT5G24770 AT5G44420 AT1G70560 AT1G23320 AT4G24670 AT2G23170 AT1G04240 AT1G04240 AT4G02980 AT3G51800 AT2G45210
Sequence (F) atgggtccttcagagagtcct ccaggtcgcaagctcttaaa tgccattggagattctaagagg ttagttaattccaacgatgcagaa tcatttgccctgaggtacatg cttcatgttacgctcaatgcc ttttcgcaaagctcatcctaa tccagtctctacctcaacacca agaagacgcgaatccagatg cactcctcaaaccccagaaa tgcccacaagctctctgata tccggaagcttttgttcaat cgttcatggttgtggtgttg cgcttaagaacaggattcagtg ggaaagcaggttggatgatct tgaactttacggcgcagact cgtcttgtagtggaagcaatga cgtggatttacgatgatttgaa tgaaatggaggagttctttgc ccctaggagtaatttggattcg aaagtgagattgaagacttctttgc gaaaccgaaaccgaaacctc gcaggcagagcttgatgact tcggtttctttacgctcgtc caagcaatctttcagggagact cgcgtgtctctcattagctg gccagcacaaccaaaggtat ttggcctttttcattagtctcttt gaaagggaggcaagggatta ttatacgcatgagattgttactctttg cacagtggaaaccatcgactc aaacgtgcgcctatggttac cttacccgcggatctcatc caccggcgttagtcaaatct gcgttgaacggagtaccaa ggttcgttctacggacgttt cgtcgattcgaaaaccatct gagttgtgcgagaagccaagt agcagagctggagagcgttgtg cttcatgttggcgagtctctcgag aaggttgtgtcagacagttgtggg catcacagagttcctcacaagc aaaggctcagattgttggatggc aagctaccaggatctttctgatgc ccaatctttgccaacagtacaa gtggtggaaagaagaagaaagc agccgaaaaagagtacggatt
Sequence (R) tggaacaccttggtcctaaag catggtagtaagtgtgatggtttcc gttgtctcggatgaaacttgc caacagactcagctggatcaaa tactccacaaccactggcaac gcaagtctagcccgaaacact cctttgaggagaattgaatggt cctttttgctccctcgaaa atctgacaatcttcgatttctgg cgagaaacatgacgagcaga tcaattgtagaagccgtttctct tgttaatccaggcatgagtcc cgcctttctctttcgtgaag ctcagctgcgaggttttgta ccgtcataaacatgtaccttctaca cggaacccacgtttgtagtatt cgacgtttcagtgtcaccat gcggcgagactctacatctt gggggatatcattcacaatgtc cactttcagaaacagaatcagacc ttgtccgagacaatgtcaaagt ccctcactcactggactcgt tcattgacgatgtcgtagttgtact aaggaacgagaatttcctcca attgggtgtcccaaacaact catactttgtctcctccacttgaa cccattggg tgtttgtgaac tggtttaaactcgaggaaactga cacgtaataccttacggtgtcg ctccgaaccatctcagcaa ggctcgtactgcagcatttta tcgacgctgtcatcttaacg actccatgttctgcggtctt ccggcggattctaagaaaa caggcacatgtgggaacc cctcacgacatggaacctg ggcaccgtgtcgaagtctat gttgcatgatccatgtttgg cttcatgttggcgagtctctcgag acgctggtcagagttatgagacacc ggttgtggctcaaagaccctgc gtcggtccatgtcttcatca tgaccctcatgctcagattcattc attccttgtgctccatagtttccc ctcatgaccggtgtttttga gcattactacttgcgtccattg gacccgttcgaaatctgaata
Summary
Gene ID AT5G53300 AT5G14520 AT2G40360 AT5G15550 AT1G07370 AT2G31270 AT3G54710 AT2G29680 AT4G14700 AT4G12620 AT2G37560 AT3G23580 AT2G36010 AT5G22220 AT1G47870 AT3G12280 AT2G23430 AT3G50630 AT5G48820 AT2G32710 AT3G24810 AT3G19150 AT1G49620 AT1G70210 AT1G76540 AT4g34160 AT5G65420 AT4G03270 AT1G07820 AT3G54180 AT1G76540 AT2G44810 AT3G45140 AT5G42650 AT2G06050 AT1G19640 AT5G24770 AT5G44420 AT1G70560 AT1G23320 AT4G24670 AT2G23170 AT1G04240 AT1G04240 AT4G02980 AT3G51800 AT2G45210 Sequence (F) atgggtccttcagagagtcct ccaggtcgcaagctcttaaa tgccattggagattctaagagg ttagttaattccaacgatgcagaa tcatttgccctgaggtacatg cttcatgttacgctcaatgcc ttttcgcaaagctcatcctaa tccagtctctacctcaacacca agaagacgcgaatccagatg cactcctcaaaccccagaaa tgcccacaagctctctgata tccggaagcttttgttcaat cgttcatggttgtggtgttg cgcttaagaacaggattcagtg ggaaagcaggttggatgatct tgaactttacggcgcagact cgtcttgtagtggaagcaatga cgtggatttacgatgatttgaa tgaaatggaggagttctttgc ccctaggagtaatttggattcg aaagtgagattgaagacttctttgc gaaaccgaaaccgaaacctc gcaggcagagcttgatgact tcggtttctttacgctcgtc caagcaatctttcagggagact cgcgtgtctctcattagctg gccagcacaaccaaaggtat ttggcctttttcattagtctcttt gaaagggaggcaagggatta ttatacgcatgagattgttactctttg cacagtggaaaccatcgactc aaacgtgcgcctatggttac cttacccgcggatctcatc caccggcgttagtcaaatct gcgttgaacggagtaccaa ggttcgttctacggacgttt cgtcgattcgaaaaccatct gagttgtgcgagaagccaagt agcagagctggagagcgttgtg cttcatgttggcgagtctctcgag aaggttgtgtcagacagttgtggg catcacagagttcctcacaagc aaaggctcagattgttggatggc aagctaccaggatctttctgatgc ccaatctttgccaacagtacaa gtggtggaaagaagaagaaagc agccgaaaaagagtacggatt
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