Internucleosomal DNA fragmentation in wild emmer wheat is catalyzed by S1-type endonucleases translocated to the nucleus upon induction of cell death

Gila Granot, Yaakov Morgenstern, Asif Khan, Yemima Givaty Rapp, Anat Pesok, Eviatar Nevo, Gideon Grafi

פרסום מחקרי: פרסום בכתב עתמאמרביקורת עמיתים

תקציר

Leaves of cereal plants display nucleosomal fragmentation of DNA attributed to the action of nucleases induced during program cell death (PCD). Yet, the specific nuclease activity responsible for generating double strand DNA breaks (DSBs) that lead to DNA fragmentation has not been fully described. Here, we characterized a Ca2+/Mg2+-dependent S1-type endonuclease activity in leaves of wild emmer wheat (Triticum dicoccoides Köern.) capable of introducing DSBs as demonstrated by the conversion of supercoiled plasmid DNA into a linear duplex DNA. In-gel nuclease assay revealed a nuclease of about 35kDa capable of degrading both single stranded DNA and RNA. We further showed that the endonuclease activity can be purified on Concanavalin A and treatment with peptide-N-glycosidase F (PNGase F) did not abolish its activity. Furthermore, ConA-associated endonuclease was capable of generating nucleosomal DNA fragmentation in tobacco nuclei. Since S1-type endonucleases lack canonical nuclear localization signal it was necessary to determine their subcellular localization. To this end, a cDNA encoding for a putative 34kDa S1-type nuclease, designated TaS1-like (TaS1L) was synthesized based on available sequence data of Triticum aestivum and fused with RFP. Introduction into protoplasts showed that TaS1L-RFP is cytoplasmic 24h post transformation but gradually turn nuclear at 48h concomitantly with induction of cell death. Our results suggest that DNA fragmentation occurring in leaves of wild emmer wheat may be attributed to S1-type endonuclease(s) that reside in the cytoplasm but translocate to the nucleus upon induction of cell death.

שפה מקוריתאנגלית אמריקאית
עמודים (מ-עד)239-246
מספר עמודים8
כתב עתBiochimica et Biophysica Acta - Gene Regulatory Mechanisms
כרך1849
מספר גיליון3
מזהי עצם דיגיטלי (DOIs)
סטטוס פרסוםפורסם - 1 מרץ 2015

ASJC Scopus subject areas

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  • ???subjectarea.asjc.1300.1304???
  • ???subjectarea.asjc.1300.1315???
  • ???subjectarea.asjc.1300.1303???

טביעת אצבע

להלן מוצגים תחומי המחקר של הפרסום 'Internucleosomal DNA fragmentation in wild emmer wheat is catalyzed by S1-type endonucleases translocated to the nucleus upon induction of cell death'. יחד הם יוצרים טביעת אצבע ייחודית.

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