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Title: Phosphorylation of spore coat proteins by a family of atypical protein kinases

Abstract

The modification of proteins by phosphorylation occurs in all life forms and is catalyzed by a large superfamily of enzymes known as protein kinases. We recently discovered a family of secretory pathway kinases that phosphorylate extracellular proteins. One member, family with sequence similarity 20C (Fam20C), is the physiological Golgi casein kinase. While examining distantly related protein sequences, we observed low levels of identity between the spore coat protein H (CotH), and the Fam20C-related secretory pathway kinases. CotH is a component of the spore in many bacterial and eukaryotic species, and is required for efficient germination of spores in Bacillus subtilis; however, the mechanism by which CotH affects germination is unclear. In this paper, we show that CotH is a protein kinase. The crystal structure of CotH reveals an atypical protein kinase-like fold with a unique mode of ATP binding. Examination of the genes neighboring cotH in B. subtilis led us to identify two spore coat proteins, CotB and CotG, as CotH substrates. Furthermore, we show that CotH-dependent phosphorylation of CotB and CotG is required for the efficient germination of B. subtilis spores. Finally and collectively, our results define a family of atypical protein kinases and reveal an unexpected role formore » protein phosphorylation in spore biology.« less

Authors:
 [1];  [2];  [1];  [3];  [4];  [5];  [6];  [2];  [3];  [7];  [5];  [8];  [2]
  1. Univ. of California, San Diego, CA (United States). Dept. of Pharmacology
  2. Univ. of Texas Southwestern Medical Center, Dallas, TX (United States). Dept. of Molecular Biology
  3. Univ. of California, San Diego, CA (United States). Division of Biological Sciences
  4. Polish Academy of Sciences (PAS), Warsaw (Poland). Inst. of Biochemistry and Biophysics. Dept. of Microbial Biochemistry
  5. Warsaw Univ. of Life Sciences (Poland). Dept. of Experimental Design and Bioinformatics
  6. Polish Academy of Sciences (PAS), Warsaw (Poland). Inst. of Biochemistry and Biophysics. Dept. of Biophysics
  7. Univ. of Texas Southwestern Medical Center, Dallas, TX (United States). Dept. of Biophysics and Biochemistry
  8. Univ. of California, San Diego, CA (United States). Dept. of Pharmacology. Dept. of Cellular and Molecular Medicine. Dept. of Chemistry and Biochemistry
Publication Date:
Research Org.:
Univ. of California, San Diego, CA (United States); Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Inst. of Health (NIH) (United States); Polish National Science Centre (Poland)
Contributing Org.:
Polish Academy of Sciences (PAS), Warsaw (Poland); Warsaw Univ. of Life Sciences (Poland)
OSTI Identifier:
1274751
Grant/Contract Number:  
AC02-06CH11357; DK018849; DK018024; R00DK099254; GM57045; 2012/05/B/NZ3/00413; 2012/07/D/NZ2/04286; RR150033
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 113; Journal Issue: 25; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; kinase; CotH; phosphorylation; CotG; CotB

Citation Formats

Nguyen, Kim B., Sreelatha, Anju, Durrant, Eric S., Lopez-Garrido, Javier, Muszewska, Anna, Dudkiewicz, Małgorzata, Grynberg, Marcin, Yee, Samantha, Pogliano, Kit, Tomchick, Diana R., Pawłowski, Krzysztof, Dixon, Jack E., and Tagliabracci, Vincent S. Phosphorylation of spore coat proteins by a family of atypical protein kinases. United States: N. p., 2016. Web. doi:10.1073/pnas.1605917113.
Nguyen, Kim B., Sreelatha, Anju, Durrant, Eric S., Lopez-Garrido, Javier, Muszewska, Anna, Dudkiewicz, Małgorzata, Grynberg, Marcin, Yee, Samantha, Pogliano, Kit, Tomchick, Diana R., Pawłowski, Krzysztof, Dixon, Jack E., & Tagliabracci, Vincent S. Phosphorylation of spore coat proteins by a family of atypical protein kinases. United States. doi:10.1073/pnas.1605917113.
Nguyen, Kim B., Sreelatha, Anju, Durrant, Eric S., Lopez-Garrido, Javier, Muszewska, Anna, Dudkiewicz, Małgorzata, Grynberg, Marcin, Yee, Samantha, Pogliano, Kit, Tomchick, Diana R., Pawłowski, Krzysztof, Dixon, Jack E., and Tagliabracci, Vincent S. Mon . "Phosphorylation of spore coat proteins by a family of atypical protein kinases". United States. doi:10.1073/pnas.1605917113. https://www.osti.gov/servlets/purl/1274751.
@article{osti_1274751,
title = {Phosphorylation of spore coat proteins by a family of atypical protein kinases},
author = {Nguyen, Kim B. and Sreelatha, Anju and Durrant, Eric S. and Lopez-Garrido, Javier and Muszewska, Anna and Dudkiewicz, Małgorzata and Grynberg, Marcin and Yee, Samantha and Pogliano, Kit and Tomchick, Diana R. and Pawłowski, Krzysztof and Dixon, Jack E. and Tagliabracci, Vincent S.},
abstractNote = {The modification of proteins by phosphorylation occurs in all life forms and is catalyzed by a large superfamily of enzymes known as protein kinases. We recently discovered a family of secretory pathway kinases that phosphorylate extracellular proteins. One member, family with sequence similarity 20C (Fam20C), is the physiological Golgi casein kinase. While examining distantly related protein sequences, we observed low levels of identity between the spore coat protein H (CotH), and the Fam20C-related secretory pathway kinases. CotH is a component of the spore in many bacterial and eukaryotic species, and is required for efficient germination of spores in Bacillus subtilis; however, the mechanism by which CotH affects germination is unclear. In this paper, we show that CotH is a protein kinase. The crystal structure of CotH reveals an atypical protein kinase-like fold with a unique mode of ATP binding. Examination of the genes neighboring cotH in B. subtilis led us to identify two spore coat proteins, CotB and CotG, as CotH substrates. Furthermore, we show that CotH-dependent phosphorylation of CotB and CotG is required for the efficient germination of B. subtilis spores. Finally and collectively, our results define a family of atypical protein kinases and reveal an unexpected role for protein phosphorylation in spore biology.},
doi = {10.1073/pnas.1605917113},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 25,
volume = 113,
place = {United States},
year = {2016},
month = {5}
}

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