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MiPRISMA: Interactome profiling of small peptides and young microproteins translated from human short open reading frames Part2

ABSTRACT: All species continuously evolve small open reading frames (sORFs) that can be templated for protein synthesis and may provide raw material for evolutionary adaptation. We analyzed the evolutionary origins of 7,264 recently cataloged human sORFs and found that most were evolutionary young and emerged de novo. We additionally discovered 221 yet uncataloged peptides smaller than 16 amino acids, for which we found evidence of translation in human and rodent tissues. To investigate the potential bioactivity of microproteins and peptides translated from young and very small ORFs we established MiPRISMA: a mass spectrometry-based interactome screen with sequence motif resolution. We assessed 266 candidates and implicated several in essential cellular processes including splicing, translational regulation, and endocytosis, a subset of which we validate in cellular assays. MiPRISMA provides a scalable platform to characterize small and evolutionarily young proteins, shedding light on a largely unexplored territory of the putative human proteome.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Marieluise Kirchner

LAB HEAD: Philipp Mertins

PROVIDER: PXD033629 | Pride | 2023-02-17

REPOSITORIES: Pride

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			Action	DRS
	ANAKIN_20191105_MK_PRISMA_2_D_001.raw	Raw
	ANAKIN_20191105_MK_PRISMA_2_D_002.raw	Raw
	ANAKIN_20191105_MK_PRISMA_2_D_003.raw	Raw
	ANAKIN_20191105_MK_PRISMA_2_D_004.raw	Raw
	ANAKIN_20191105_MK_PRISMA_2_D_005.raw	Raw

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Project description:The development of transcriptomic tools has allowed exhaustive description of stress responses. These responses always superimpose a general response associated to growth rate decrease and a specific one corresponding to the stress. The exclusive growth rate response can be achieved through chemostat cultivation, enabling all parameters to remain constant except the growth rate. We analysed metabolic and transcriptomic responses of Lactococcus lactis in continuous cultures at different growth rates ranging from 0.09 to 0.47 h-1. Growth rate was conditioned by isoleucine supply. Although the metabolism was constant and homolactic, a widespread transcriptomic response involving 30 % of the genome was observed. The expression of genes encoding physiological functions associated with biogenesis increased with growth rate (transcription, translation, fatty acid and phospholipids metabolism). Many phages, prophages and transposons related genes were down regulated by growth rate suggesting genome plasticity to be involved in the adaptation to slow growth. The growth rate response was compared to carbon and amino-acid starvation transcriptomic responses, revealing constant and significant involvement of growth rate regulations in these two stressful conditions (overlap 26%). Although stringent response mechanism is considered as the one governing growth deceleration in bacteria, the rigorous comparison of the two transcriptomic responses clearly indicated the mechanisms are distinct. Moreover it was established that genes positively regulated by growth rate are preferentially located in the vicinity of replication origin while those negatively regulated are mainly encountered at the opposite. This result demonstrates the often neglected relationship between genes expression and their location on chromosome. Keywords: growth rate impact, continuous cultures Continuous cultivation of Lactococcus lactis IL1403 were carried out on a chemically defined medium and under controlled conditions (30 Â°C, pH 6.6, nitrogen atmosphere). Cell samples were harvested at steady state. Total RNA was extracted from these samples and radiolabelled cDNA were prepared and hybridized on nylon arrays. 1948 amplicons specific of Lactococcus lactis IL1403 genes were spotted twice on the array. Samples corresponding to various growth rates were analyzed simultaneously and 3 independent repetitions were performed.

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MiPRISMA: Interactome profiling of small peptides and young microproteins translated from human short open reading frames Part2

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