The efficacy of current antimalarial drugs is threatened by reduced susceptibility of Plasmodium falciparum to artemisinin, associated with mutations in pfkelch13 Another gene with variants known to modulate the response to artemisinin encodes the μ subunit of the AP-2 adaptin trafficking complex. To elucidate the cellular role of AP-2μ in P. falciparum, we performed a conditional gene knockout, which severely disrupted schizont organization and maturation, leading to mislocalization of key merozoite proteins. AP-2μ is thus essential for blood-stage replication. We generated transgenic P. falciparum parasites expressing hemagglutinin-tagged AP-2μ and examined cellular localization by fluorescence and electron microscopy. Together with mass spectrometry analysis of coimmunoprecipitating proteins, these studies identified AP-2μ-interacting partners, including other AP-2 subunits, the K10 kelch-domain protein, and PfEHD, an effector of endocytosis and lipid mobilization, but no evidence was found of interaction with clathrin, the expected coat protein for AP-2 vesicles. In reverse immunoprecipitation experiments with a clathrin nanobody, other heterotetrameric AP-complexes were shown to interact with clathrin, but AP-2 complex subunits were absent.IMPORTANCE We examine in detail the AP-2 adaptin complex from the malaria parasite Plasmodium falciparum In most studied organisms, AP-2 is involved in bringing material into the cell from outside, a process called endocytosis. Previous work shows that changes to the μ subunit of AP-2 can contribute to drug resistance. Our experiments show that AP-2 is essential for parasite development in blood but does not have any role in clathrin-mediated endocytosis. This suggests that a specialized function for AP-2 has developed in malaria parasites, and this may be important for understanding its impact on drug resistance.

• MeSH
• adaptorový proteinový komplex 2 genetika   metabolismus   MeSH
• antimalarika farmakologie   MeSH
• artemisininy metabolismus   MeSH
• endocytóza fyziologie   MeSH
• geneticky modifikované organismy MeSH
• genový knockout MeSH
• klathrin metabolismus   MeSH
• léková rezistence MeSH
• membránové proteiny metabolismus   MeSH
• Plasmodium falciparum účinky léků   genetika   metabolismus   MeSH
• protozoální proteiny genetika   metabolismus   MeSH
• schizonti účinky léků   genetika   metabolismus   MeSH
• transport proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Jasmonates (JAs) are signals in plant stress responses and development. One of the first observed and prominent responses to JAs is the induction of biosynthesis of different groups of secondary compounds. Among them are nicotine, isoquinolines, glucosinolates, anthocyanins, benzophenanthridine alkaloids, artemisinin, and terpenoid indole alkaloids (TIAs), such as vinblastine. This brief review describes modes of action of JAs in the biosynthesis of anthocyanins, nicotine, TIAs, glucosinolates and artemisinin. After introducing JA biosynthesis, the central role of the SCFCOI1-JAZ co-receptor complex in JA perception and MYB-type and MYC-type transcription factors is described. Brief comments are provided on primary metabolites as precursors of secondary compounds. Pathways for the biosynthesis of anthocyanin, nicotine, TIAs, glucosinolates and artemisinin are described with an emphasis on JA-dependent transcription factors, which activate or repress the expression of essential genes encoding enzymes in the biosynthesis of these secondary compounds. Applied aspects are discussed using the biotechnological formation of artemisinin as an example of JA-induced biosynthesis of secondary compounds in plant cell factories.

• MeSH
• anthokyaniny biosyntéza   MeSH
• artemisininy metabolismus   MeSH
• biologické modely MeSH
• biosyntetické dráhy MeSH
• cyklopentany metabolismus   MeSH
• glukosinoláty biosyntéza   MeSH
• metabolické inženýrství MeSH
• nikotin biosyntéza   MeSH
• oxylipiny metabolismus   MeSH
• regulátory růstu rostlin biosyntéza   metabolismus   MeSH
• rostlinné proteiny metabolismus   MeSH
• rostliny genetika   metabolismus   MeSH
• sekologanin-tryptaminové alkaloidy metabolismus   MeSH
• signální transdukce MeSH
• transkripční faktory metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Plant secondary metabolism evolved in the context of highly organized and differentiated cells and tissues, featuring massive chemical complexity operating under tight environmental, developmental and genetic control. Biotechnological demand for natural products has been continuously increasing because of their significant value and new applications, mainly as pharmaceuticals. Aseptic production systems of plant secondary metabolites have improved considerably, constituting an attractive tool for increased, stable and large-scale supply of valuable molecules. Surprisingly, to date, only a few examples including taxol, shikonin, berberine and artemisinin have emerged as success cases of commercial production using this strategy. The present review focuses on the main characteristics of plant specialized metabolism and their implications for current strategies used to produce secondary compounds in axenic cultivation systems. The search for consonance between plant secondary metabolism unique features and various in vitro culture systems, including cell, tissue, organ, and engineered cultures, as well as heterologous expression in microbial platforms, is discussed. Data to date strongly suggest that attaining full potential of these biotechnology production strategies requires being able to take advantage of plant specialized metabolism singularities for improved target molecule yields and for bypassing inherent difficulties in its rational manipulation.

• MeSH
• artemisininy izolace a purifikace   metabolismus   MeSH
• axenická kultura MeSH
• berberin izolace a purifikace   metabolismus   MeSH
• biologické přípravky izolace a purifikace   metabolismus   MeSH
• biotechnologie metody   MeSH
• buněčné kultury MeSH
• fytonutrienty biosyntéza   izolace a purifikace   MeSH
• metabolické inženýrství metody   MeSH
• naftochinony izolace a purifikace   metabolismus   MeSH
• paclitaxel biosyntéza   izolace a purifikace   MeSH
• rostlinné buňky chemie   metabolismus   MeSH
• rostliny chemie   genetika   metabolismus   MeSH
• sekundární metabolismus MeSH
• techniky tkáňových kultur MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH