N-Terminal Lipid Modification Is Required for the Stable Accumulation of CyanoQ in Synechocystis sp. PCC 6803
Abstract
The CyanoQ protein has been demonstrated to be a component of cyanobacterial Photosystem II (PS II), but there exist a number of outstanding questions concerning its physical association with the complex. CyanoQ is a lipoprotein; upon cleavage of its transit peptide by Signal Peptidase II, which targets delivery of the mature protein to the thylakoid lumenal space, the N-terminal cysteinyl residue is lipid-modified. This modification appears to tether this otherwise soluble component to the thylakoid membrane. To probe the functional significance of the lipid anchor, mutants of the CyanoQ protein have been generated in Synechocystis sp. PCC 6803 to eliminate the N-terminal cysteinyl residue, preventing lipid modification. Substitution of the N-terminal cysteinyl residue with serine (Q-C22S) resulted in a decrease in the amount of detectable CyanoQ protein to 17% that of the wild-type protein. Moreover, the physical properties of the accumulated Q-C22S protein were consistent with altered processing of the CyanoQ precursor. The Q-C22S protein was shifted to a higher apparent molecular mass and partitioned in the hydrophobic phase in TX-114 phase-partitioning experiments. These results suggest that the hydrophobic N-terminal 22 amino acids were not properly cleaved by a signal peptidase. Substitution of the entire CyanoQ transit peptide with themore »
- Authors:
- Publication Date:
- Research Org.:
- Louisiana State Univ., Baton Rouge, LA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1337766
- Alternate Identifier(s):
- OSTI ID: 1361695; OSTI ID: 1393395
- Grant/Contract Number:
- FG02-09ER20310; FG02-98ER20310; FG02-09ER16070; SC0002628
- Resource Type:
- Published Article
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Name: PLoS ONE Journal Volume: 11 Journal Issue: 9; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science (PLoS)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; lipids; signal peptides; peripheral membrane proteins; synechocystis; signal processing; proteases; membrane proteins; outer membrane proteins; 60 APPLIED LIFE SCIENCES; Peripheral Membranes; Synchocystis
Citation Formats
Juneau, Andrea D., Frankel, Laurie K., Bricker, Terry M., Roose, Johnna L., and Theg, ed., Steven M. N-Terminal Lipid Modification Is Required for the Stable Accumulation of CyanoQ in Synechocystis sp. PCC 6803. United States: N. p., 2016.
Web. doi:10.1371/journal.pone.0163646.
Juneau, Andrea D., Frankel, Laurie K., Bricker, Terry M., Roose, Johnna L., & Theg, ed., Steven M. N-Terminal Lipid Modification Is Required for the Stable Accumulation of CyanoQ in Synechocystis sp. PCC 6803. United States. https://doi.org/10.1371/journal.pone.0163646
Juneau, Andrea D., Frankel, Laurie K., Bricker, Terry M., Roose, Johnna L., and Theg, ed., Steven M. Thu .
"N-Terminal Lipid Modification Is Required for the Stable Accumulation of CyanoQ in Synechocystis sp. PCC 6803". United States. https://doi.org/10.1371/journal.pone.0163646.
@article{osti_1337766,
title = {N-Terminal Lipid Modification Is Required for the Stable Accumulation of CyanoQ in Synechocystis sp. PCC 6803},
author = {Juneau, Andrea D. and Frankel, Laurie K. and Bricker, Terry M. and Roose, Johnna L. and Theg, ed., Steven M.},
abstractNote = {The CyanoQ protein has been demonstrated to be a component of cyanobacterial Photosystem II (PS II), but there exist a number of outstanding questions concerning its physical association with the complex. CyanoQ is a lipoprotein; upon cleavage of its transit peptide by Signal Peptidase II, which targets delivery of the mature protein to the thylakoid lumenal space, the N-terminal cysteinyl residue is lipid-modified. This modification appears to tether this otherwise soluble component to the thylakoid membrane. To probe the functional significance of the lipid anchor, mutants of the CyanoQ protein have been generated in Synechocystis sp. PCC 6803 to eliminate the N-terminal cysteinyl residue, preventing lipid modification. Substitution of the N-terminal cysteinyl residue with serine (Q-C22S) resulted in a decrease in the amount of detectable CyanoQ protein to 17% that of the wild-type protein. Moreover, the physical properties of the accumulated Q-C22S protein were consistent with altered processing of the CyanoQ precursor. The Q-C22S protein was shifted to a higher apparent molecular mass and partitioned in the hydrophobic phase in TX-114 phase-partitioning experiments. These results suggest that the hydrophobic N-terminal 22 amino acids were not properly cleaved by a signal peptidase. Substitution of the entire CyanoQ transit peptide with the transit peptide of the soluble lumenal protein PsbO yielded the Q-SS mutant and resulted in no detectable accumulation of the modified CyanoQ protein. Finally, the CyanoQ protein was present at normal amounts in the PS II mutant strains ΔpsbB and ΔpsbO, indicating that an association with PS II was not a prerequisite for stable CyanoQ accumulation. Together these results indicate that CyanoQ accumulation in Synechocystis sp. PCC 6803 depends on the presence of the N-terminal lipid anchor, but not on the association of CyanoQ with the PS II complex.},
doi = {10.1371/journal.pone.0163646},
journal = {PLoS ONE},
number = 9,
volume = 11,
place = {United States},
year = {Thu Sep 22 00:00:00 EDT 2016},
month = {Thu Sep 22 00:00:00 EDT 2016}
}
https://doi.org/10.1371/journal.pone.0163646
Web of Science
Works referenced in this record:
[34] Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes
book, January 1987
- Lichtenthaler, Hartmut K.
- Methods in Enzymology
Crystal structure of CyanoQ from the thermophilic cyanobacterium Thermosynechococcus elongatus and detection in isolated photosystem II complexes
journal, May 2014
- Michoux, Franck; Boehm, Marko; Bialek, Wojciech
- Photosynthesis Research, Vol. 122, Issue 1
Homologs of Plant PsbP and PsbQ Proteins Are Necessary for Regulation of Photosystem II Activity in the Cyanobacterium Synechocystis 6803
journal, July 2004
- Thornton, Leeann E.; Ohkawa, Hiroshi; Roose, Johnna L.
- The Plant Cell, Vol. 16, Issue 8
Post-translational protein translocation into thylakoids by the Sec and ΔpH-dependent pathways
journal, December 2001
- Mori, Hiroki; Cline, Kenneth
- Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1541, Issue 1-2
Regulation of Manganese Uptake in Synechocystis 6803 by RfrA, a Member of a Novel Family of Proteins Containing a Repeated Five-Residues Domain †
journal, May 2003
- Chandler, Leeann E.; Bartsevich, Victor V.; Pakrasi, Himadri B.
- Biochemistry, Vol. 42, Issue 18
Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of thylakoid membranes at 4 C: Characterizations of two additional chlorophyll a-protein complexes
journal, January 1979
- Delepelaire, P.; Chua, N. -H.
- Proceedings of the National Academy of Sciences, Vol. 76, Issue 1
Photosystem II characteristics of a constructedSynechocystis 6803 mutant lacking synthesis of the D1 polypeptide
journal, June 1990
- Nilsson, Fredrik; Andersson, Bertil; Jansson, Christer
- Plant Molecular Biology, Vol. 14, Issue 6
Absence of the PsbQ Protein Results in Destabilization of the PsbV Protein and Decreased Oxygen Evolution Activity in Cyanobacterial Photosystem II
journal, May 2006
- Kashino, Yasuhiro; Inoue-Kashino, Natsuko; Roose, Johnna L.
- Journal of Biological Chemistry, Vol. 281, Issue 30
Native structure of photosystem II at 1.95 Å resolution viewed by femtosecond X-ray pulses
journal, November 2014
- Suga, Michihiro; Akita, Fusamichi; Hirata, Kunio
- Nature, Vol. 517, Issue 7532
Triton X-114 phase fractionation of membrane proteins of the cyanobacterium Anacystis nidulans R2
journal, November 1984
- Bricker, Terry M.; Sherman, Louis A.
- Archives of Biochemistry and Biophysics, Vol. 235, Issue 1
Evidence that D1 Processing Is Required for Manganese Binding and Extrinsic Protein Assembly into Photosystem II
journal, August 2004
- Roose, Johnna L.; Pakrasi, Himadri B.
- Journal of Biological Chemistry, Vol. 279, Issue 44
Multiple copies of the PsbQ protein in a cyanobacterial photosystem II assembly intermediate complex
journal, March 2015
- Liu, Haijun; Weisz, Daniel A.; Pakrasi, Himadri B.
- Photosynthesis Research, Vol. 126, Issue 2-3
Triton X-114 phase-fractionation of maize thylakoid membranes in the investigation of thylakoid protein topology
journal, November 1982
- Bricker, Terry M.; Sherman, Louis A.
- FEBS Letters, Vol. 149, Issue 2
The extrinsic proteins of photosystem II: update
journal, January 2016
- Roose, Johnna L.; Frankel, Laurie K.; Mummadisetti, Manjula P.
- Planta, Vol. 243, Issue 4
Sorting of lipoproteins to the outer membrane in E. coli
journal, July 2004
- Tokuda, Hajime; Matsuyama, Shin-ichi
- Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1693, Issue 1
A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K
journal, January 1988
- Taylor, Laura A.; Rose, Ronald E.
- Nucleic Acids Research, Vol. 16, Issue 15
The extrinsic proteins of Photosystem II
journal, January 2012
- Bricker, Terry M.; Roose, Johnna L.; Fagerlund, Robert D.
- Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1817, Issue 1
SignalP 4.0: discriminating signal peptides from transmembrane regions
journal, September 2011
- Petersen, Thomas Nordahl; Brunak, Søren; von Heijne, Gunnar
- Nature Methods, Vol. 8, Issue 10
Physiologically active chloroplasts contain pools of unassembled extrinsic proteins of the photosynthetic oxygen-evolving enzyme complex in the thylakoid lumen
journal, October 1991
- Ettinger, W. F.
- The Journal of Cell Biology, Vol. 115, Issue 2
Crystal structure of oxygen-evolving photosystem II at a resolution of 1.9 Å
journal, April 2011
- Umena, Yasufumi; Kawakami, Keisuke; Shen, Jian-Ren
- Nature, Vol. 473, Issue 7345
Prediction of lipoprotein signal peptides in Gram-negative bacteria
journal, August 2003
- Juncker, Agnieszka S.; Willenbrock, Hanni; von Heijne, Gunnar
- Protein Science, Vol. 12, Issue 8
The PsbP and PsbQ family proteins in the photosynthetic machinery of chloroplasts
journal, August 2014
- Ifuku, Kentaro
- Plant Physiology and Biochemistry, Vol. 81
Site-directed mutagenesis of the CPa-1 protein of photosystem II: Alteration of the basic residue pair 384,385R to 384,385G leads to a defect associated with the oxygen-evolving complex
journal, November 1992
- Putnam-Evans, Cindy; Bricker, Terry M.
- Biochemistry, Vol. 31, Issue 46
The Sll0606 Protein Is Required for Photosystem II Assembly/Stability in the Cyanobacterium Synechocystis sp. PCC 6803
journal, August 2010
- Zhang, Shulu; Frankel, Laurie K.; Bricker, Terry M.
- Journal of Biological Chemistry, Vol. 285, Issue 42
MS-based cross-linking analysis reveals the location of the PsbQ protein in cyanobacterial photosystem II
journal, February 2014
- Liu, H.; Zhang, H.; Weisz, D. A.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 12
Lipoproteins in bacteria
journal, June 1990
- Hayashi, Shigeru; Wu, Henry C.
- Journal of Bioenergetics and Biomembranes, Vol. 22, Issue 3
The Bacterial Twin-Arginine Translocation Pathway
journal, October 2006
- Lee, Philip A.; Tullman-Ercek, Danielle; Georgiou, George
- Annual Review of Microbiology, Vol. 60, Issue 1
NIH Image to ImageJ: 25 years of image analysis
journal, June 2012
- Schneider, Caroline A.; Rasband, Wayne S.; Eliceiri, Kevin W.
- Nature Methods, Vol. 9, Issue 7
PsbQ (Sll1638) in Synechocystis sp. PCC 6803 Is Required for Photosystem II Activity in Specific Mutants and in Nutrient-Limiting Conditions †
journal, January 2005
- Summerfield, Tina C.; Shand, Jackie A.; Bentley, Fiona K.
- Biochemistry, Vol. 44, Issue 2
The PsbQ protein defines cyanobacterial Photosystem II complexes with highest activity and stability
journal, February 2007
- Roose, J. L.; Kashino, Y.; Pakrasi, H. B.
- Proceedings of the National Academy of Sciences, Vol. 104, Issue 7
Lipoprotein Sorting in Bacteria
journal, October 2011
- Okuda, Suguru; Tokuda, Hajime
- Annual Review of Microbiology, Vol. 65, Issue 1
A method for analyzing lipid-modified proteins with mass spectrometry
journal, March 2008
- Ujihara, Tetsuro; Sakurai, Isamu; Mizusawa, Naoki
- Analytical Biochemistry, Vol. 374, Issue 2