A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs
The slow kinetics and poor substrate specificity of the key photosynthetic CO2-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as pyrenoids in the majority of eukaryotic microalgae. Diatoms dominate marine photosynthesis, but the intera...
| Main Authors: | , , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
| Published: |
2024
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| Online Access: | https://hdl.handle.net/10356/174561 |
| _version_ | 1826111440695590912 |
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| author | Oh, Zhen Guo Ang, Warren Shou Leong Poh, Cheng Wei Lai, Soak-Kuan Sze, Siu Kwan Li, Hoi-Yeung Bhushan, Shashi Wunder, Tobias Mueller-Cajar, Oliver |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Oh, Zhen Guo Ang, Warren Shou Leong Poh, Cheng Wei Lai, Soak-Kuan Sze, Siu Kwan Li, Hoi-Yeung Bhushan, Shashi Wunder, Tobias Mueller-Cajar, Oliver |
| author_sort | Oh, Zhen Guo |
| collection | NTU |
| description | The slow kinetics and poor substrate specificity of the key photosynthetic CO2-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as pyrenoids in the majority of eukaryotic microalgae. Diatoms dominate marine photosynthesis, but the interactions underlying their pyrenoids are unknown. Here, we identify and characterize the Rubisco linker protein PYCO1 from Phaeodactylum tricornutum. PYCO1 is a tandem repeat protein containing prion-like domains that localizes to the pyrenoid. It undergoes homotypic liquid-liquid phase separation (LLPS) to form condensates that specifically partition diatom Rubisco. Saturation of PYCO1 condensates with Rubisco greatly reduces the mobility of droplet components. Cryo-electron microscopy and mutagenesis data revealed the sticker motifs required for homotypic and heterotypic phase separation. Our data indicate that the PYCO1-Rubisco network is cross-linked by PYCO1 stickers that oligomerize to bind to the small subunits lining the central solvent channel of the Rubisco holoenzyme. A second sticker motif binds to the large subunit. Pyrenoidal Rubisco condensates are highly diverse and tractable models of functional LLPS. |
| first_indexed | 2024-10-01T02:50:45Z |
| format | Journal Article |
| id | ntu-10356/174561 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:50:45Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | ntu-10356/1745612024-08-07T02:32:29Z A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs Oh, Zhen Guo Ang, Warren Shou Leong Poh, Cheng Wei Lai, Soak-Kuan Sze, Siu Kwan Li, Hoi-Yeung Bhushan, Shashi Wunder, Tobias Mueller-Cajar, Oliver School of Biological Sciences Nanyang Institute of Structural Biology Medicine, Health and Life Sciences Pyrenoid Rubisco The slow kinetics and poor substrate specificity of the key photosynthetic CO2-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as pyrenoids in the majority of eukaryotic microalgae. Diatoms dominate marine photosynthesis, but the interactions underlying their pyrenoids are unknown. Here, we identify and characterize the Rubisco linker protein PYCO1 from Phaeodactylum tricornutum. PYCO1 is a tandem repeat protein containing prion-like domains that localizes to the pyrenoid. It undergoes homotypic liquid-liquid phase separation (LLPS) to form condensates that specifically partition diatom Rubisco. Saturation of PYCO1 condensates with Rubisco greatly reduces the mobility of droplet components. Cryo-electron microscopy and mutagenesis data revealed the sticker motifs required for homotypic and heterotypic phase separation. Our data indicate that the PYCO1-Rubisco network is cross-linked by PYCO1 stickers that oligomerize to bind to the small subunits lining the central solvent channel of the Rubisco holoenzyme. A second sticker motif binds to the large subunit. Pyrenoidal Rubisco condensates are highly diverse and tractable models of functional LLPS. Published version The research was supported by Ministry of Education of Singapore grants MOE2018-T2-2-059 and MOE2019-T3-1-012 (to O.M.-C). 2024-04-02T06:56:42Z 2024-04-02T06:56:42Z 2023 Journal Article Oh, Z. G., Ang, W. S. L., Poh, C. W., Lai, S., Sze, S. K., Li, H., Bhushan, S., Wunder, T. & Mueller-Cajar, O. (2023). A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs. Proceedings of the National Academy of Sciences, 120(25), e2304833120-. https://dx.doi.org/10.1073/pnas.2304833120 0027-8424 https://hdl.handle.net/10356/174561 10.1073/pnas.2304833120 37311001 2-s2.0-85161909179 25 120 e2304833120 en MOE2018-T2-2-059 MOE2019-T3-1-012 Proceedings of the National Academy of Sciences 10.21979/N9/IXBAW5 © 2023 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY). application/pdf |
| spellingShingle | Medicine, Health and Life Sciences Pyrenoid Rubisco Oh, Zhen Guo Ang, Warren Shou Leong Poh, Cheng Wei Lai, Soak-Kuan Sze, Siu Kwan Li, Hoi-Yeung Bhushan, Shashi Wunder, Tobias Mueller-Cajar, Oliver A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title | A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title_full | A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title_fullStr | A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title_full_unstemmed | A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title_short | A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs |
| title_sort | linker protein from a red type pyrenoid phase separates with rubisco via oligomerizing sticker motifs |
| topic | Medicine, Health and Life Sciences Pyrenoid Rubisco |
| url | https://hdl.handle.net/10356/174561 |
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