Abstract
Key message
Homologous genes for the peptidoglycan precursor flippase MurJ, and peptidoglycan hydrolases: lytic transglycosylase MltB, and dd-carboxypeptidase VanY are required for chloroplast division in the moss Physcomitrella patens.
Abstract
The moss Physcomitrella patens is used as a model plant to study plastid peptidoglycan biosynthesis. In bacteria, MurJ flippase transports peptidoglycan precursors from the cytoplasm to the periplasm. In this study, we identified a MurJ homolog (PpMurJ) in the P. patens genome. Bacteria employ peptidoglycan degradation and recycling pathways for cell division. We also searched the P. patens genome for genes homologous to bacterial peptidoglycan hydrolases and identified genes homologous for the lytic transglycosylase mltB, N-acetylglucosaminidase nagZ, and ld-carboxypeptidase ldcA in addition to a putative dd-carboxypeptidase vanY reported previously. Moreover, we found a ß-lactamase-like gene (Pplactamase). GFP fusion proteins with either PpMltB or PpVanY were detected in the chloroplasts, whereas fusion proteins with PpNagZ, PpLdcA, or Pplactamase localized in the cytoplasm. Experiments seeking PpMurJ-GFP fusion proteins failed. PpMurJ gene disruption in P. patens resulted in the appearance of macrochloroplasts in protonemal cells. Compared with the numbers of chloroplasts in wild-type plants (38.9 ± 4.9), PpMltB knockout and PpVanY knockout had lower numbers of chloroplasts (14.3 ± 6.7 and 28.1 ± 5.9, respectively). No differences in chloroplast numbers were observed after PpNagZ, PpLdcA, or Pplactamase single-knockout. Chloroplast numbers in PpMltB/PpVanY double-knockout cells were similar to those in PpMltB single-knockout cells. Zymogram analysis of the recombinant PpMltB protein revealed its peptidoglycan hydrolase activity. Our results imply that PpMurJ, PpMltB and PpVanY play a critical role in chloroplast division in the moss P. patens.
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Acknowledgements
This work was supported by the Japan Society for the Promotion of Science KAKENHI (Grant Number 17H03701). We are grateful to Sayaka Koshigae, and Ayane Tanaka (Kumamoto University) for their assistance.
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The Japan Society for the Promotion of Science KAKENHI (Grant Number 17H03701).
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HT and KT designed the study, analyzed the data, and wrote the manuscript. HU, NS, HK, MF, SH, MU and KT performed the experiments. All authors read and approved the final manuscript.
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Utsunomiya, H., Saiki, N., Kadoguchi, H. et al. Genes encoding lipid II flippase MurJ and peptidoglycan hydrolases are required for chloroplast division in the moss Physcomitrella patens. Plant Mol Biol 107, 405–415 (2021). https://doi.org/10.1007/s11103-020-01081-0
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DOI: https://doi.org/10.1007/s11103-020-01081-0