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Medium-scale Preparation of Drosophila Embryo Extracts for Proteomic Experiments
Published on: May 30, 2017
Systematic Identification of Microproteins during the Development of Drosophila melanogaster
Zhiwei Wang1, Ni Pan1, Jiahao Yan1
1School of Life Sciences and Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, Wuhan, Hubei 430079, People's Republic of China.
Abstract:
Short open reading frame-encoded peptides (SEPs) are microproteins with less than 100 amino acids that play an essential role in the growth and development of organisms. There are plenty of short open reading frames in Drosophila melanogaster that potentially code polypeptides. We chose 11 time points during the life cycle of Drosophila to investigate microproteins, particularly those related to development. Finally, we identified a total of 410 microproteins, of which 27 were noncoding RNA-encoded proteins. Of the 410 microproteins, 74 were expressed in all stages from embryo to adults, whereas 300 microproteins were only found in one or two time points. Approximately, one-third of the microproteins were not reported previously and 44 were obtained from de novo sequencing, validated by synthetic peptides. These microproteins are related to the main bioprocesses of growth and development, such as multicellular organism reproduction, postmating behavior, and oviposition. Over half of the microproteins have predicted functional domains and are conserved across species, suggesting that these microproteins have critical functions in fly development. This work enriches the D. melanogaster proteome and provides a significant data resource for growth and development research.
Insights
Researchers identified 410 short open reading frame-encoded peptides (SEPs) in Drosophila melanogaster, revealing novel microproteins crucial for organismal growth and development. Many SEPs are conserved and functionally important, enriching the fly proteome.
Area of Science:
- * Molecular Biology
- * Developmental Biology
- * Genomics
Background:
- * Short open reading frame-encoded peptides (SEPs) are microproteins (<100 amino acids) vital for organismal growth and development.
- * The Drosophila melanogaster genome contains numerous short open reading frames with potential to encode polypeptides.
- * Understanding SEP roles is crucial for comprehending fundamental biological processes.
Purpose of the Study:
- * To investigate microproteins, particularly those involved in development, across 11 life cycle time points in Drosophila melanogaster.
- * To identify and characterize novel SEPs and assess their conservation and potential functions.
- * To expand the known proteome of Drosophila melanogaster and provide a resource for developmental research.
Main Methods:
- * Analysis of 11 distinct developmental time points in the life cycle of Drosophila melanogaster.
- * Identification of microproteins encoded by short open reading frames.
- * De novo sequencing and synthetic peptide validation for novel microproteins.
- * Prediction of functional domains and cross-species conservation analysis.
Main Results:
- * A total of 410 microproteins were identified, including 27 noncoding RNA-encoded proteins.
- * 74 microproteins were ubiquitously expressed, while 300 were stage-specific.
- * Approximately one-third of identified microproteins were previously unreported, with 44 validated by de novo sequencing.
- * Over half of the microproteins possess predicted functional domains and are conserved across species.
- * Identified microproteins are linked to key developmental processes like reproduction and oviposition.
Conclusions:
- * This study significantly expands the known Drosophila melanogaster proteome by identifying 410 microproteins.
- * A substantial portion of these microproteins are novel, conserved, and functionally relevant to development.
- * The findings provide a valuable data resource for future research into growth and developmental mechanisms in flies and other organisms.
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