Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling
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- dc.contributor.author Hong, Shin-Young
- dc.contributor.author Sun, Bin
- dc.contributor.author Straub, Daniel
- dc.contributor.author Blaakmeer, Anko
- dc.contributor.author Mineri, Lorenzo
- dc.contributor.author Koch, Jonas
- dc.contributor.author Brinch-Pedersen, Henrik
- dc.contributor.author Holme, Inger B.
- dc.contributor.author Burow, Meike
- dc.contributor.author Lyngs Jørgensen, Hans Jørgen
- dc.contributor.author Albà Soler, Mar
- dc.contributor.author Wenkel, Stephan
- dc.date.accessioned 2021-03-24T06:57:56Z
- dc.date.available 2021-03-24T06:57:56Z
- dc.date.issued 2020
- dc.description.abstract MicroProteins are small, often single-domain proteins that are sequence-related to larger, often multidomain proteins. Here, we used a combination of comparative genomics and heterologous synthetic misexpression to isolate functional cereal microProtein regulators. Our approach identified LITTLE NINJA (LNJ), a microProtein that acts as a modulator of jasmonic acid (JA) signaling. Ectopic expression of LNJ in Arabidopsis resulted in stunted plants that resembled the decuple JAZ (jazD) mutant. In fact, comparing the transcriptomes of transgenic LNJ overexpressor plants and jazD revealed a large overlap of deregulated genes, suggesting that ectopic LNJ expression altered JA signaling. Transgenic Brachypodium plants with elevated LNJ expression levels showed deregulation of JA signaling as well and displayed reduced growth and enhanced production of side shoots (tiller). This tillering effect was transferable between grass species, and overexpression of LNJ in barley and rice caused similar traits. We used a clustered regularly interspaced short palindromic repeats (CRISPR) approach and created a LNJ-like protein in Arabidopsis by deleting parts of the coding sentence of the AFP2 gene that encodes a NINJA-domain protein. These afp2-crispr mutants were also stunted in size and resembled jazD Thus, similar genome-engineering approaches can be exploited as a future tool to create LNJ proteins and produce cereals with altered architectures.
- dc.description.sponsorship Work in the laboratory was funded by the European Research Council (ERC-StG miPDesign, 336295), the Independent Research Fund Denmark (DFF-FNU, Adaptogenomics), the NovoNordisk Foundation (NNF18OC0034226), and start-up funding of Copenhagen Plant Science Centre through the University of Copenhagen. We thank the anonymous reviewers from a previous submission, Hans Thordal-Christensen, Gregg Howe, and Michael Broberg Palmgren for critically reading and improving our manuscript.
- dc.format.mimetype application/pdf
- dc.identifier.citation Hong SY, Sun B, Straub D, Blaakmeer A, Mineri L, Koch J, Brinch-Pedersen H, Holme IB, Burow M, Lyngs Jørgensen HJ, Albà MM, Wenkel S. Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling. Proc Natl Acad Sci U S A. 2020; 117(42):26197-205. DOI: 10.1073/pnas.2005198117
- dc.identifier.doi http://dx.doi.org/10.1073/pnas.2005198117
- dc.identifier.issn 0027-8424
- dc.identifier.uri http://hdl.handle.net/10230/46911
- dc.language.iso eng
- dc.publisher National Academy of Sciences
- dc.relation.ispartof Proc Natl Acad Sci U S A. 2020; 117(42):26197-205
- dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/336295
- dc.rights © 2020 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).
- dc.rights.accessRights info:eu-repo/semantics/openAccess
- dc.rights.uri http://creativecommons.org/licenses/by/4.0/
- dc.subject.keyword Branching
- dc.subject.keyword Jasmonic acid
- dc.subject.keyword microProteins
- dc.title Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling
- dc.type info:eu-repo/semantics/article
- dc.type.version info:eu-repo/semantics/publishedVersion