Intein Inhibitors as Novel Antimicrobials: Protein Splicing in Human Pathogens, Screening Methods, and Off-Target Considerations

Wall, Diana A.; Tarrant, Seanan P.; Wang, Qianqian; Mills, Kenneth V.; Lennon, Christopher

doi:10.3389/fmolb.2021.752824

Cited by 8 publications

(4 citation statements)

References 104 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2) Transesterification : The hydroxyl group of cholesterol non-covalently bound to the sterol recognition region (SRR) of HhC carries out a nucleophilic attack on the thioester, releasing HhN and linking it covalently to cholesterol ( Ciulla et al, 2019 ). Inteins, on the other hand, catalyze protein splicing processes in four steps ( Wall et al, 2021 ), with the first two steps very similar to Hh autoprocessing ( Koonin, 1995 ; Hall et al, 1997 ; Pietrokovski, 1998 ), except that the intramolecular transesterification in intein splicing is replaced by an intermolecular transesterification, namely cholesteroylation of HhN in Hh proteins ( Porter et al, 1996b ). While Hh Hint and inteins have conserved C258 and TXXH motif in common, Hh Hint has unique residues not present in inteins, e.g., D303 and C400 Drosophila Hint domain.…”

Section: Structural Mechanism Of Hedgehog Autoprocessingmentioning

confidence: 99%

Hedgehog Autoprocessing: From Structural Mechanisms to Drug Discovery

Kandel

Wang

2022

Front. Mol. Biosci.

Self Cite

View full text Add to dashboard Cite

Hedgehog (Hh) signaling plays pivotal roles in embryonic development. In adults, Hh signaling is mostly turned off but its abnormal activation is involved in many types of cancer. Hh signaling is initiated by the Hh ligand, generated from the Hh precursor by a specialized autocatalytic process called Hh autoprocessing. The Hh precursor consists of an N-terminal signaling domain (HhN) and a C-terminal autoprocessing domain (HhC). During Hh autoprocessing, the precursor is cleaved between N- and C-terminal domain followed by the covalent ligation of cholesterol to the last residue of HhN, which subsequently leads to the generation of Hh ligand for Hh signaling. Hh autoprocessing is at the origin of canonical Hh signaling and precedes all downstream signaling events. Mutations in the catalytic residues in HhC can lead to congenital defects such as holoprosencephaly (HPE). The aim of this review is to provide an in-depth summary of the progresses and challenges towards an atomic level understanding of the structural mechanisms of Hh autoprocessing. We also discuss drug discovery efforts to inhibit Hh autoprocessing as a new direction in cancer therapy.

show abstract

Section: Structural Mechanism Of Hedgehog Autoprocessingmentioning

confidence: 99%

Hedgehog Autoprocessing: From Structural Mechanisms to Drug Discovery

Kandel

Wang

2022

Front. Mol. Biosci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…[ 35 , 36 , 37 , 38 , 39 , 40 ]. The inhibition of Prp8 intein splicing, resulting in a non-functional host protein, has been proposed as a new possible approach for antifungal treatment [ 41 , 42 , 43 ]. In vivo and also in vitro screening systems for such inhibition were created mainly for RecA intein in an attempt to screen possible antituberculosis compounds [ 44 , 45 , 46 , 47 ] and, recently, an in vitro screening system based on green fluorescent protein was applied for evaluating the inhibition of Prp8 intein from C. neoformans and C. gattii (CnePrp8i and CgaPrp8i, respectively) by compounds from a small-molecule library [ 48 ], however, still, in vivo systems for evaluating CnePrp8i splicing have yet to be developed.…”

Section: Introductionmentioning

confidence: 99%

Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics

Fernandes

Zatti²,

Arantes³

et al. 2022

JoF

View full text Add to dashboard Cite

Inteins are genetic mobile elements that are inserted within protein-coding genes, which are usually housekeeping genes. They are transcribed and translated along with the host gene, then catalyze their own splicing out of the host protein, which assumes its functional conformation thereafter. As Prp8 inteins are found in several important fungal pathogens and are absent in mammals, they are considered potential therapeutic targets since inhibiting their splicing would selectively block the maturation of fungal proteins. We developed a target-based drug screening system to evaluate the splicing of Prp8 intein from the yeast pathogen Cryptococcus neoformans (CnePrp8i) using Saccharomyces cerevisiae Ura3 as a non-native host protein. In our heterologous system, intein splicing preserved the full functionality of Ura3. To validate the system for drug screening, we examined cisplatin, which has been described as an intein splicing inhibitor. By using our system, new potential protein splicing inhibitors may be identified and used, in the future, as a new class of drugs for mycosis treatment. Our system also greatly facilitates the visualization of CnePrp8i splicing dynamics in vivo.

show abstract

“…Mycobacterium tuberculosis and Cryptococcus neoformans, they are absent from the genomes of humans. Inteins therefore represent attractive antimicrobial targets, whereby the inhibition of splicing could compromise pathogen survival(Wall et al, 2021;Tharappel et al, 2022). Excitingly, recent work has demonstrated that small molecules can target protein splicing as an antimicrobial strategy(Chan et al, 2016;Li et al, 2019Li et al, , 2021.…”

mentioning

confidence: 99%

Inteins—mechanism of protein splicing, emerging regulatory roles, and applications in protein engineering

Wood,

Belfort,

Lennon

2023

Front. Microbiol.

Self Cite

View full text Add to dashboard Cite

Protein splicing is a posttranslational process in which an intein segment excises itself from two flanking peptides, referred to as exteins. In the native context, protein splicing results in two separate protein products coupled to the activation of the intein-containing host protein. Inteins are generally described as either full-length inteins, mini-inteins or split inteins, which are differentiated by their genetic structure and features. Inteins can also be divided into three classes based on their splicing mechanisms, which differ in the location of conserved residues that mediate the splicing pathway. Although inteins were once thought to be selfish genetic elements, recent evidence suggests that inteins may confer a genetic advantage to their host cells through posttranslational regulation of their host proteins. Finally, the ability of modified inteins to splice and cleave their fused exteins has enabled many new applications in protein science and synthetic biology. In this review, we briefly cover the mechanisms of protein splicing, evidence for some inteins as environmental sensors, and intein-based applications in protein engineering.

show abstract

Intein Inhibitors as Novel Antimicrobials: Protein Splicing in Human Pathogens, Screening Methods, and Off-Target Considerations

Cited by 8 publications

References 104 publications

Hedgehog Autoprocessing: From Structural Mechanisms to Drug Discovery

Hedgehog Autoprocessing: From Structural Mechanisms to Drug Discovery

Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics

Inteins—mechanism of protein splicing, emerging regulatory roles, and applications in protein engineering

Contact Info

Product

Resources

About