Vpliv proteinov NLP na kvasovke

magistrsko delo

Anja Porenta (Author), Uroš Petrovič (Reviewer), Kristina Sepčić (Mentor), Apolonija Bedina Zavec (Co-mentor)

Abstract

Proteine iz družine NLP (Nep1-like proteins) najdemo v raznolikih, taksonomsko nesorodnih mikroorganizmih (glivah, oomicetah, bakterijah). Takšni mikroorganizmi so zelo razširjeni in lahko okužijo različne pridelke, predvsem dvokaličnice, kot so krompir, paradižnik, soja, tobak in hmelj. Nekateri proteini NLP so citotoksični za dvokaličnice saj povzročijo nekrozo tkiva in aktivirajo imunski odziv rastline. Poleg citotoksičnih proteinov pa obstajajo tudi proteini NLP, ki niso citotoksični. Proteini NLP so po strukturi podobni aktinoporinom, toksinom, ki tvorijo pore v celično membrano; in se prav tako kot aktinoporini vežejo na sfingolipide. Za proučevanje interakcije med proteini NLP in sfingolipidi smo izbrali kvasovko Saccharomyces cerevisiae, ki je enostaven in dobro poznan modelni organizem ter hkrati vsebuje sfingolipide, ki so v precejšnji meri podobni rastlinskim sfingolipidom. Ugotovili smo, da se citotoksični protein NLPPya veže na kvasne protoplaste močneje in je tudi bolj toksičen za kvasovke od proteina HaNLP3, ki ni cititoksičen za rastline. Za proučevanje vezave proteinov NLP na kvasne sfingolipide smo uporabili dva mutirana seva kvasovke, ki imata okvarjeno manozilacijo sfingolipidov. Ugotovili smo, da se največ proteina veže na sev BY4742 csg2Δ, ki nima manoziliranih sfingolipidov, ima pa povečano količino inozitol-fosforil ceramidov.

Keywords

proteini Nep 1-like;proteini NLP;kvasni sfingolipidi;kvasovke;citotoksičnost;vezava;kvasni protoplasti;NLPpya;HaNLP3;

Data

Language:	Slovenian
Year of publishing:	2018
Typology:	2.09 - Master's Thesis
Organization:	UL BF - Biotechnical Faculty
Publisher:	[A. Porenta]
UDC:	577.112+577.2:582.282.23
COBISS:	5010296
Views:	720
Downloads:	251
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Effect of NLP proteins on yeasts
Secondary abstract:	Proteins from the NLP family (Nep1-like proteins) are found in a variety of taxonomically unrelated microorganisms (fungi, oomycetes, bacteria). Such microorganisms are widespread and can infect different crop plants, especially dicotyledons such as potatoe, tomatoe, soybean, tobacco and hop. Some NLP proteins are cytotoxic to dicotyledonous plants, and cause tissue necrosis and induce immune response of the plant. In addition to cytotoxic proteins, there are also NLP proteins that are not cytotoxic. NLP proteins are structurally similar to actinoporins, toxins that form pores into the cell membrane. NLP proteins bind to sphingolipids, just like actinoporins. The yeast Saccharomyces cerevisiae, which is a simple and well-known model organism and contains sphingolipids that are similar to plant sphingolipids, has been selected for the study of the interaction between NLP proteins and sphingolipids. We found that the cytotoxic protein NLPPya binds to the yeast protoplasts more strongly and is also more toxic to yeasts than the protein HaNLP3, which is not cytotoxic to plants. To examine the binding of NLP proteins to yeast sphingolipids we used two mutants with impaired sphingolipid mannosylation, and compared them to the wild type cells. We found that most proteins bind to the BY4742 csg2Δ strain which does not have manosylated sphingolipids, but has an increased amount of ceramides and inositol-phosphoryl ceramides.
Secondary keywords:	Nep 1-like proteins;NLP proteins;yeast sphingolipids;yeasts;cytotoxicity;binding;yeast protoplasts;NLPpya;HaNLP3;
Type (COBISS):	Master's thesis/paper
Study programme:	0
Thesis comment:	Univ. v Ljubljani, Biotehniška fak., Študij mikrobiologije
Pages:	XI, 72 f.
ID:	10994682