Dataset Information

Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein.

ABSTRACT: Intein-mediated protein splicing has been widely used in protein engineering; however, the splicing efficiency and extein specificity usually limit its further application. Thus, there is a demand for more general inteins that can overcome these limitations. Here, we study the trans-splicing of CPE intein obtained from the directed evolution of Cne PRP8, which shows that its splicing rate is ~29- fold higher than that of the wild-type. When the +1 residue of C-extein is changed to cysteine, CPE also shows high splicing activity. Faster association and higher affinity may contribute to the high splicing rate compared with wild-type intein. These findings have important implications for the future engineering of inteins and provide clues for fundamental studies of protein structure and folding.

SUBMITTER: Zhan Q

PROVIDER: S-EPMC10448054 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein.

Zhan Qin Q Shi Changhua C Jiang Yu Y Gao Xianling X Lin Ying Y

Acta biochimica et biophysica Sinica 20230701 8

Intein-mediated protein splicing has been widely used in protein engineering; however, the splicing efficiency and extein specificity usually limit its further application. Thus, there is a demand for more general inteins that can overcome these limitations. Here, we study the trans-splicing of CPE intein obtained from the directed evolution of Cne PRP8, which shows that its splicing rate is ~29- fold higher than that of the wild-type. When the +1 residue of C-extein is chan ...[more]

PMID: 37489009

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Project description:BackgroundWe recently described a mini-intein in the PRP8 gene of a strain of the basidiomycete Cryptococcus neoformans, an important fungal pathogen of humans. This was the second described intein in the nuclear genome of any eukaryote; the first nuclear encoded intein was found in the VMA gene of several saccharomycete yeasts. The evolution of eukaryote inteins is not well understood. In this report we describe additional PRP8 inteins (bringing the total of these to over 20). We compare and contrast the phylogenetic distribution and evolutionary history of the PRP8 intein and the saccharomycete VMA intein, in order to derive a broader understanding of eukaryote intein evolution. It has been suggested that eukaryote inteins undergo horizontal transfer and the present analysis explores this proposal.ResultsIn total, 22 PRP8 inteins have been detected in species from three different orders of euascomycetes, including Aspergillus nidulans and Aspergillus fumigatus (Eurotiales), Paracoccidiodes brasiliensis, Uncinocarpus reesii and Histoplasma capsulatum (Onygales) and Botrytis cinerea (Helotiales). These inteins are all at the same site in the PRP8 sequence as the original Cryptococcus neoformans intein. Some of the PRP8 inteins contain apparently intact homing endonuclease domains and are thus potentially mobile, while some lack the region corresponding to the homing endonuclease and are thus mini-inteins. In contrast, no mini-inteins have been reported in the VMA gene of yeast. There are several examples of pairs of closely related species where one species carries the PRP8 intein while the intein is absent from the other species. Bio-informatic and phylogenetic analyses suggest that many of the ascomycete PRP8 homing endonucleases are active. This contrasts with the VMA homing endonucleases, most of which are inactive.ConclusionPRP8 inteins are widespread in the euascomycetes (Pezizomycota) and apparently their homing endonucleases are active. There is no evidence for horizontal transfer within the euascomycetes. This suggests that the intein is of ancient origin and has been vertically transmitted amongst the euascomycetes. It is possible that horizontal transfer has occurred between the euascomycetes and members of the basidiomycete genus Cryptococcus.

Dataset Information

Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein.

Publications

Efficient splicing of the CPE intein derived from directed evolution of the <i>Cryptococcus neoformans</i> PRP8 intein.

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Dataset Information

Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein.

Publications

Efficient splicing of the CPE intein derived from directed evolution of the &lt;i&gt;Cryptococcus neoformans&lt;/i&gt; PRP8 intein.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Efficient splicing of the CPE intein derived from directed evolution of the <i>Cryptococcus neoformans</i> PRP8 intein.