Award Abstract # 1745036
Collaborative Research: Antarctic Diatom Proteorhodopsins: Characterization and a Potential Role in the Iron-limitation Response

NSF Org: OPP
Office of Polar Programs (OPP)
Recipient: UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL
Initial Amendment Date: May 1, 2018
Latest Amendment Date: May 19, 2021
Award Number: 1745036
Award Instrument: Standard Grant
Program Manager: Rebecca Gast
rgast@nsf.gov
 (703)292-2356
OPP
 Office of Polar Programs (OPP)
GEO
 Directorate for Geosciences
Start Date: June 1, 2018
End Date: September 30, 2023 (Estimated)
Total Intended Award Amount: $511,278.00
Total Awarded Amount to Date: $606,742.00
Funds Obligated to Date: FY 2018 = $511,278.00
FY 2021 = $95,464.00
History of Investigator:
  • Adrian Marchetti (Principal Investigator)
    amarchetti@unc.edu
  • Alecia Septer (Co-Principal Investigator)
Recipient Sponsored Research Office: University of North Carolina at Chapel Hill
104 AIRPORT DR STE 2200
CHAPEL HILL
NC  US  27599-5023
(919)966-3411
Sponsor Congressional District: 04
Primary Place of Performance: University of North Carolina at Chapel Hill
Chapel Hill
NC  US  27599-1350
Primary Place of Performance
Congressional District:
04
Unique Entity Identifier (UEI): D3LHU66KBLD5
Parent UEI: D3LHU66KBLD5
NSF Program(s): ANT Organisms & Ecosystems
Primary Program Source: 0100XXXXDB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s): 097Z
Program Element Code(s): 511100
Award Agency Code: 4900
Fund Agency Code: 4900
Assistance Listing Number(s): 47.078

ABSTRACT

Proteorhodopsins are proteins that are embedded in membranes that can act as light-driven proton pumps to generate energy for metabolism and growth. The discovery of proteorhodopsins in many diverse marine prokaryotic microbes has initiated extensive investigation into their distributions and functional roles. Recently, a proton-pumping, rhodopsin-like gene was identified in diatoms, a group of marine phytoplankton that dominates the base of the food web in much of the Southern Ocean. Since this time, proteorhodopsins have been identified in many, but not all, diatom species. The proteorhodopsin gene is more frequently found in diatoms residing in cold, iron-limited regions of the ocean, including the Southern Ocean, than in diatoms from other regions. It is thought that proteorhodopsin is especially suited for use energy production in the Southern Ocean since it uses no iron and its reaction rate is insensitive to temperature (unlike conventional photosynthesis). The overall objective of the project is to characterize Antarctic diatom-proteorhodopsin and determine its role in the adaptation of these diatoms to low iron concentrations and extremely low temperatures found in Antarctic waters. This research will provide new information on the genetic underpinnings that contribute to the success of diatoms in the Southern Ocean and how this unique molecule may play a pivotal role in providing energy to the base of the Antarctic food web. Broader impact activities are aimed to promote the teaching and learning of polar marine-sciences related topics by translating research objectives into readily accessible educational materials for middle-school students.

This project will combine molecular, biochemical and physiological measurements to determine the role and importance of proteorhodopsin in diatom isolates from the Western Antarctic Peninsula region. Proton-pumping characteristics and pumping rates of proteorhodopsin as a function of light intensity and temperature, the resultant proteorhodopsin-linked intracellular ATP production rates, and the cellular localization of the protein will be determined. The project will examine the environmental conditions where Antarctic diatom-proteorhodopsin is most highly expressed and construct a cellular energy budget that includes diatom-proteorhodopsin when grown under these different environmental conditions. Estimates of the energy flux generated by proteorhodopsin will be compared to total energy generation by the photosynthetic light reactions and metabolically coupled respiration rates. Finally, the characteristics and gene expression of diatom-proteorhodopsin in Antarctic diatoms and a proteorhodopsin-containing diatom isolates from temperate regions will be compared in order to determine if there is a preferential dependence on energy production through proteorhodopsin in diatoms residing in cold, iron-limited regions of the ocean. Educational activities will be performed in collaboration with the Morehead Planetarium and Science Center who co-ordinates the SciVentures program, a popular summer camp for middle-school students from Chapel Hill and surrounding areas. In collaboration with the Planetarium, the researchers will develop activities that focus on phytoplankton and the important role they play within polar marine food webs for the SciVentures participants. Additionally, a teaching module on Antarctic phytoplankton will be developed for classrooms and made available to educational networking websites and presented at workshops for science educators nationwide.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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Andrew, Sarah and Wilson, Travis and Smith, Stephanie and Marchetti, Adrian and Septer, Alecia N. "A tripartite model system for Southern Ocean diatom-bacterial interactions reveals the coexistence of competing symbiotic strategies" ISME Communications , v.2 , 2022 https://doi.org/10.1038/s43705-022-00181-w Citation Details
Andrew, Sarah M. and Moreno, Carly M. and Plumb, Kaylie and Hassanzadeh, Babak and Gomez-Consarnau, Laura and Smith, Stephanie N. and Schofield, Oscar and Yoshizawa, Susumu and Fujiwara, Takayoshi and Sunda, William G. and Hopkinson, Brian M. and Septer, "Widespread use of proton-pumping rhodopsin in Antarctic phytoplankton" Proceedings of the National Academy of Sciences , v.120 , 2023 https://doi.org/10.1073/pnas.2307638120 Citation Details
Brown, Michael S. and Bowman, Jeff S. and Lin, Yajuan and Feehan, Colette J. and Moreno, Carly M. and Cassar, Nicolas and Marchetti, Adrian and Schofield, Oscar M. "Low diversity of a key phytoplankton group along the West Antarctic Peninsula" Limnology and Oceanography , v.66 , 2021 https://doi.org/10.1002/lno.11765 Citation Details
Lin, Yajuan and Moreno, Carly and Marchetti, Adrian and Ducklow, Hugh and Schofield, Oscar and Delage, Erwan and Meredith, Michael and Li, Zuchuan and Eveillard, Damien and Chaffron, Samuel and Cassar, Nicolas "Decline in plankton diversity and carbon flux with reduced sea ice extent along the Western Antarctic Peninsula" Nature Communications , v.12 , 2021 https://doi.org/10.1038/s41467-021-25235-w Citation Details
Moreno, Carly M. and Gong, Weida and Cohen, Natalie R. and DeLong, Kimberly and Marchetti, Adrian "Interactive effects of iron and light limitation on the molecular physiology of the Southern Ocean diatom Fragilariopsiskerguelensis" Limnology and Oceanography , v.65 , 2020 https://doi.org/10.1002/lno.11404 Citation Details
Yoshizawa, Susumu and Azuma, Tomonori and Kojima, Keiichi and Inomura, Keisuke and Hasegawa, Masumi and Nishimura, Yosuke and Kikuchi, Masuzu and Armin, Gabrielle and Tsukamoto, Yuya and Miyashita, Hideaki and Ifuku, Kentaro and Yamano, Takashi and Marche "Light-driven Proton Pumps as a Potential Regulator for Carbon Fixation in Marine Diatoms" Microbes and Environments , v.38 , 2023 https://doi.org/10.1264/jsme2.ME23015 Citation Details

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

Proton-pumping rhodopsins (PPRs) are retinal-binding membrane proteins that function as light-driven proton pumps that create proton gradients across cell membranes, which can be used for active membrane transport, and potentially, the production of ATP (adenosine triphosphate), the energy currency of all cells needed for metabolism and growth. Unlike photosynthesis which is heavily dependent on the micronutrient iron, PPR is iron-free, and thus, should be favored in low-iron ocean waters such as those that occur in the Southern Ocean (SO). In this study, we showed that PPR in the Southern Ocean diatom isolate Pseudo-nitzschia subcurvata is localized to the vacuole, making it a possible alternative phototrophic organelle for light-driven production of cellular energy.  Estimates of energy flux through PPR by way of retinal quantification in three SO diatoms suggest that PPR may produce a significant amount of cellular energy under low temperature, low iron and high light conditions, as commonly found in surface waters of the Southern Ocean.  Through a combination of gene expression analysis and retinal quantification in samples collected during a field survey of marine waters surrounding the West Antarctic Peninsula, we show that use of PPR is widespread in this polar environment and increases in iron-limited regions.  Taken together these findings point to PPRs being an important adaptation of SO phytoplankton for their growth and survival, and thus, may have a significant impact on the global carbon cycling.

A post-doc, three graduate students, and several undergraduate students were trained as part of this project.  Polar diatoms isolated as part of this project have been further distributed to other members of the scientific community to support their research.  This grant also supported development of methods for the cultivation and examination of diatom-associated bacteria as well as an immunofluorescence  microscopy protocol to localize proteins of interest, relative to other cellular structures, within a silicious polar diatom. Activities to teach elementary students about polar marine-sciences related topics were created through translating our research objectives into readily accessible educational materials during several community engagement events.  Data associated with this project is made available at the publication websites and USAP-DC.


Last Modified: 12/30/2023
Modified by: Adrian Marchetti

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