Progeny of the flesh fly Sarcophaga bullata exposed to short day length show a maternal effect that prevents the expression of pupal diapause. Although ecological aspects of this effect are well studied, not enough is known about the molecular mechanisms underlying this maternal effect. In this study, two-dimensional electrophoresis was performed to detect differences of the abundance of certain proteins in the ovaries of this fly kept under long day and short day conditions for 2 days after eclosion. Eleven proteins that were abundant and showed significant changes were selected for mass spectrometric identification. Ovary proteins that increased in abundance under short-day conditions were similar to twinstar CG4254-PA, muscle protein 20-like protein, GA13413-PA, gene analogous to small peritrophins (Gasp CG10287-PA) and Ribosomal protein LP1 CG4087-PA. Ovary proteins that decreased in abundance under short-day conditions were similar to the ATP synthase beta subunit, fk506-binding protein and storage protein-binding protein. The 2-D proteome maps included 2 additional unknown proteins that were more abundant and 1 that was less abundant in the ovaries of 2-day old short-day females. Twinstar CG4254-PA, muscle protein 20-like protein and GA13413-PA harbour an actin-binding domain. That the 3 actin-binding proteins increase in abundance suggests that it is likely that an alteration in the actin cytoskeleton is involved in this maternal effect in the flesh fly., Aiqing Li ... [et al.]., and Obsahuje seznam literatury
Transcription factors exert their regulatory potential on RNA polymerase II machinery through a multiprotein complex called Mediator complex or Mediator. The Mediator complex integrates regulatory signals from cell regulatory cascades with the regulation by transcription factors. The Mediator complex consists of 25 subunits in Saccharomyces cerevisiae and 30 or more subunits in multicellular eukaryotes. Mediator subunit 28 (MED28), along with MED30, MED23, MED25 and MED26, belong to presumably evolutionarily new subunits that seem to be absent in unicellular eukaryotes and are likely to have evolved together with multicellularity and cell differentiation. Previously, we have shown that an originally uncharacterized predicted gene, F28F8.5, is the true MED28 orthologue in Caenorhabditis elegans (mdt-28) and showed that it is involved in a spectrum of developmental processes. Here, we studied the proteomic interactome of MDT-28 edited as GFP::MDT-28 using Crispr/Cas9 technology or MDT-28::GFP expressed from extrachromosomal arrays in transgenic C. elegans exploiting the GFPTRAP system and mass spectrometry. The results show that MDT-28 associates with the Head module subunits MDT-6, MDT-8, MDT-11, MDT-17, MDT20, MDT-22, and MDT-30 and the Middle module subunit MDT-14. The analyses also identified additional proteins as preferential MDT-28 interactants, including chromatin-organizing proteins, structural proteins and enzymes. The results provide evidence for MDT-28 engagement in the Mediator Head module and support the possibility of physical (direct or indirect) interaction of MDT-28 with additional proteins, reflecting the transcription-regulating potential of primarily structural and enzymatic proteins at the level of the Mediator complex.