scholarly journals Protein Splicing Activity of the Haloferax volcanii PolB-c Intein Is Sensitive to Homing Endonuclease Domain Mutations

Biochemistry ◽  
2020 ◽  
Vol 59 (36) ◽  
pp. 3359-3367 ◽  
Author(s):  
Shachar Robinzon ◽  
Alexandra R. Cawood ◽  
Mercedes A. Ruiz ◽  
Uri Gophna ◽  
Neta Altman-Price ◽  
...  
Keyword(s):  
Homing Endonuclease ◽  
Haloferax Volcanii ◽  
Protein Splicing ◽  
Endonuclease Domain

scholarly journals Algal Viruses with Distinct Intraspecies Host Specificities Include Identical Intein Elements

2005 ◽  
Vol 71 (7) ◽  
pp. 3599-3607 ◽  
Author(s):  
Keizo Nagasaki ◽  
Yoko Shirai ◽  
Yuji Tomaru ◽  
Kensho Nishida ◽  
Shmuel Pietrokovski
Keyword(s):  
Dna Polymerase ◽  
Homing Endonuclease ◽  
Sister Group ◽  
Dna Virus ◽  
Single Nucleotide Change ◽  
Double Stranded Dna ◽  
Endonuclease Domain ◽  
Algal Viruses ◽  
Molecular Phylogenetic ◽  
Position Coding

ABSTRACT Heterosigma akashiwo virus (HaV) is a large double-stranded DNA virus infecting the single-cell bloom-forming raphidophyte (golden brown alga) H. akashiwo. A molecular phylogenetic sequence analysis of HaV DNA polymerase showed that it forms a sister group with Phycodnaviridae algal viruses. All 10 examined HaV strains, which had distinct intraspecies host specificities, included an intein (protein intron) in their DNA polymerase genes. The 232-amino-acid inteins differed from each other by no more than a single nucleotide change. All inteins were present at the same conserved position, coding for an active-site motif, which also includes inteins in mimivirus (a very large double-stranded DNA virus of amoebae) and in several archaeal DNA polymerase genes. The HaV intein is closely related to the mimivirus intein, and both are apparently monophyletic to the archaeal inteins. These observations suggest the occurrence of horizontal transfers of inteins between viruses of different families and between archaea and viruses and reveal that viruses might be reservoirs and intermediates in horizontal transmissions of inteins. The homing endonuclease domain of the HaV intein alleles is mostly deleted. The mechanism keeping their sequences basically identical in HaV strains specific for different hosts is yet unknown. One possibility is that rapid and local changes in the HaV genome change its host specificity. This is the first report of inteins found in viruses infecting eukaryotic algae.

Salt‐dependent protein splicing of an intein from the halophile Haloferax volcanii

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Alexandra Cawood ◽  
Mercedes Ruiz ◽  
Kenneth Mills
Keyword(s):  
Haloferax Volcanii ◽  
Protein Splicing ◽  
Dependent Protein

Homing endonucleases residing within inteins: evolutionary puzzles awaiting genetic solutions

2011 ◽  
Vol 39 (1) ◽  
pp. 169-173 ◽  
Author(s):  
Adi Barzel ◽  
Adit Naor ◽  
Eyal Privman ◽  
Martin Kupiec ◽  
Uri Gophna
Keyword(s):  
Homing Endonuclease ◽  
Horizontal Transmission ◽  
Molecular Genetic ◽  
Halophilic Archaea ◽  
Homing Endonucleases ◽  
Protein Coding ◽  
Selfish Genetic Elements ◽  
Fitness Effects ◽  
Endonuclease Domain ◽  
Competition Assays

Inteins are selfish genetic elements that disrupt the sequence of protein-coding genes and are excised post-translationally. Most inteins also contain a HEN (homing endonuclease) domain, which is important for their horizontal transmission. The present review focuses on the evolution of inteins and their nested HENs, and highlights several unsolved questions that could benefit from molecular genetic approaches. Such approaches can be well carried out in halophilic archaea, which are naturally intein-rich and have highly developed genetic tools for their study. In particular, the fitness effects of habouring an intein/HEN can be tested in direct competition assays, providing additional insights that will improve current evolutionary models.

scholarly journals Crystal Structure of PI-SceI, a Homing Endonuclease with Protein Splicing Activity

Cell ◽  
1997 ◽  
Vol 89 (4) ◽  
pp. 555-564 ◽  
Author(s):  
Xiaoqun Duan ◽  
Frederick S Gimble ◽  
Florante A Quiocho
Keyword(s):  
Crystal Structure ◽  
Homing Endonuclease ◽  
Protein Splicing

scholarly journals Inteins in Science: Evolution to Application

2020 ◽  
Vol 8 (12) ◽  
pp. 2004
Author(s):  
Ananya Nanda ◽  
Sourya Subhra Nasker ◽  
Ashwaria Mehra ◽  
Sunita Panda ◽  
Sasmita Nayak
Keyword(s):  
Protein Modification ◽  
Genetic Manipulation ◽  
Homing Endonuclease ◽  
Linear Perspective ◽  
Endonuclease Domain ◽  
Monogenic Diseases ◽  
Genetic Mobility ◽  
Cis And Trans ◽  
And Control

Inteins are mobile genetic elements that apply standard enzymatic strategies to excise themselves post-translationally from the precursor protein via protein splicing. Since their discovery in the 1990s, recent advances in intein technology allow for them to be implemented as a modern biotechnological contrivance. Radical improvement in the structure and catalytic framework of cis- and trans-splicing inteins devised the development of engineered inteins that contribute to various efficient downstream techniques. Previous literature indicates that implementation of intein-mediated splicing has been extended to in vivo systems. Besides, the homing endonuclease domain also acts as a versatile biotechnological tool involving genetic manipulation and control of monogenic diseases. This review orients the understanding of inteins by sequentially studying the distribution and evolution pattern of intein, thereby highlighting a role in genetic mobility. Further, we include an in-depth summary of specific applications branching from protein purification using self-cleaving tags to protein modification, post-translational processing and labelling, followed by the development of intein-based biosensors. These engineered inteins offer a disruptive approach towards research avenues like biomaterial construction, metabolic engineering and synthetic biology. Therefore, this linear perspective allows for a more comprehensive understanding of intein function and its diverse applications.

scholarly journals In Vivo Characterization of the Homing Endonuclease within the polB Gene in the Halophilic Archaeon Haloferax volcanii

PLoS ONE ◽  
2011 ◽  
Vol 6 (1) ◽  
pp. e15833 ◽  
Author(s):  
Adit Naor ◽  
Rona Lazary ◽  
Adi Barzel ◽  
R. Thane Papke ◽  
Uri Gophna
Keyword(s):  
Homing Endonuclease ◽  
Haloferax Volcanii ◽  
Halophilic Archaeon ◽  
In Vivo Characterization

Homing Endonuclease and Protein Splicing Activity of Inteins from Thermophilic Archaea

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Seanan P. Tarrant ◽  
Bella J. Jankowski ◽  
Hannah Y. Comeau ◽  
Amanda C. Wibben ◽  
Kenneth V. Mills
Keyword(s):  
Homing Endonuclease ◽  
Protein Splicing ◽  
Thermophilic Archaea

scholarly journals Conditional Protein Splicing Switch in Hyperthermophiles through an Intein-Extein Partnership

mBio ◽  
2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Christopher W. Lennon ◽  
Matthew Stanger ◽  
Nilesh K. Banavali ◽  
Marlene Belfort
Keyword(s):  
Homing Endonuclease ◽  
Fine Tuning ◽  
Host Protein ◽  
Sequence Difference ◽  
Posttranslational Regulation ◽  
Protein Splicing ◽  
Microbial World ◽  
Environmental Signals ◽  
Flexible Loop ◽  
New Form

ABSTRACT Inteins are intervening proteins that undergo an autocatalytic splicing reaction that ligates flanking host protein sequences termed exteins. Some intein-containing proteins have evolved to couple splicing to environmental signals; this represents a new form of posttranslational regulation. Of particular interest is RadA from the archaeon Pyrococcus horikoshii, for which long-range intein-extein interactions block splicing, requiring temperature and single-stranded DNA (ssDNA) substrate to splice rapidly and accurately. Here, we report that splicing of the intein-containing RadA from another archaeon, Thermococcus sibericus, is activated by significantly lower temperatures than is P. horikoshii RadA, consistent with differences in their growth environments. Investigation into variations between T. sibericus and P. horikoshii RadA inteins led to the discovery that a nonconserved region (NCR) of the intein, a flexible loop where a homing endonuclease previously resided, is critical to splicing. Deletion of the NCR leads to a substantial loss in the rate and accuracy of P. horikoshii RadA splicing only within native exteins. The influence of the NCR deletion can be largely overcome by ssDNA, demonstrating that the splicing-competent conformation can be achieved. We present a model whereby the NCR is a flexible hinge which acts as a switch by controlling distant intein-extein interactions that inhibit active site assembly. These results speak to the repurposing of the vestigial endonuclease loop to control an intein-extein partnership, which ultimately allows exquisite adaptation of protein splicing upon changes in the environment. IMPORTANCE Inteins are mobile genetic elements that interrupt coding sequences (exteins) and are removed by protein splicing. They are abundant elements in microbes, and recent work has demonstrated that protein splicing can be controlled by environmental cues, including the substrate of the intein-containing protein. Here, we describe an intein-extein collaboration that controls temperature-induced splicing of RadA from two archaea and how variation in this intein-extein partnership results in fine-tuning of splicing to closely match the environment. Specifically, we found that a small sequence difference between the two inteins, a flexible loop that likely once housed a homing endonuclease used for intein mobility, acts as a switch to control intein-extein interactions that block splicing. Our results argue strongly that some inteins have evolved away from a purely parasitic lifestyle to control the activity of host proteins, representing a new form of posttranslational regulation that is potentially widespread in the microbial world.

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