Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling

Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling. Hong, S., Sun, B., Straub, D., Blaakmeer, A., Mineri, L., Koch, J., Brinch-Pedersen, H., Holme, I. B., Burow, M., Lyngs Jørgensen, H. J., Albà, M. M., & Wenkel, S. Proceedings of the National Academy of Sciences, 117(42):26197–26205, October, 2020.

Paper doi abstract bibtex

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.

@article{hong_heterologous_2020,
	title = {Heterologous {microProtein} expression identifies {LITTLE} {NINJA}, a dominant regulator of jasmonic acid signaling},
	volume = {117},
	url = {https://www.pnas.org/doi/full/10.1073/pnas.2005198117},
	doi = {10.1073/pnas.2005198117},
	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.},
	number = {42},
	urldate = {2022-11-30},
	journal = {Proceedings of the National Academy of Sciences},
	author = {Hong, Shin-Young and Sun, Bin and Straub, Daniel and Blaakmeer, Anko and Mineri, Lorenzo and Koch, Jonas and Brinch-Pedersen, Henrik and Holme, Inger B. and Burow, Meike and Lyngs Jørgensen, Hans Jørgen and Albà, M. Mar and Wenkel, Stephan},
	month = oct,
	year = {2020},
	pages = {26197--26205},
}

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