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Analysis of the Sequence Preference of Saporin by Deep Sequencing.
- Source :
-
ACS chemical biology [ACS Chem Biol] 2022 Sep 16; Vol. 17 (9), pp. 2619-2630. Date of Electronic Publication: 2022 Aug 15. - Publication Year :
- 2022
-
Abstract
- Ribosome-inactivating proteins (RIPs) are RNA:adenosine glycosidases that inactivate eukaryotic ribosomes by depurinating the sarcin-ricin loop (SRL) in 28S rRNA. The GAGA sequence at the top of the SRL or at the top of a hairpin loop is assumed to be their target motif. Saporin is a RIP widely used to develop immunotoxins for research and medical applications, but its sequence specificity has not been investigated. Here, we combine the conventional aniline cleavage assay for depurinated nucleic acids with high-throughput sequencing to study sequence-specific depurination of oligonucleotides caused by saporin. Our data reveal the sequence preference of saporin for different substrates and show that the GAGA motif is not efficiently targeted by this protein, neither in RNA nor in DNA. Instead, a preference of saporin for certain hairpin DNAs was observed. The observed sequence-specific activity of saporin may be relevant to antiviral or apoptosis-inducing effects of RIPs. The developed method could also be useful for studying the sequence specificity of depurination by other RIPs or enzymes.
- Subjects :
- Adenosine
Aniline Compounds
Antiviral Agents pharmacology
DNA metabolism
High-Throughput Nucleotide Sequencing
Oligonucleotides
Plant Proteins metabolism
RNA metabolism
RNA, Ribosomal, 28S
Ribosome Inactivating Proteins
Ribosome Inactivating Proteins, Type 1
Saporins
Immunotoxins
Ricin pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1554-8937
- Volume :
- 17
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- ACS chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 35969718
- Full Text :
- https://doi.org/10.1021/acschembio.2c00531