Crystallization and preliminary X-ray studies of I-PpoI: a nuclear, intron-encoded homing endonuclease from Physarum polycephalum

Protein Science : a Publication of the Protein Society
K E FlickB L Stoddard

Abstract

The homing endonuclease I-PpoI is encoded by an optional third intron, Pp LSU 3, found in nuclear, extrachromosomal copies of the Physarum polycephalum 26S rRNA gene. This endonuclease promotes the lateral transfer or "homing" of its encoding intron by recognizing and cleaving a partially symmetric, 15 bp homing site in 26S rDNA alleles that lack the Pp LSU 3 intron. The open reading frame encoding I-PpoI has been subcloned, and the endonuclease has been overproduced in E. coli. Purified recombinant I-PpoI has been co-crystallized with a 21 bp homing site DNA duplex. The crystals belong to space group P3(1)21, with unit cell dimensions a = b = 114 A, c = 89 A. The results of initial X-ray diffraction experiments indicate that the asymmetric unit contains an enzyme homodimer and one duplex DNA molecule, and that the unit cell has a specific volume of 3.4 A3/dalton. These experiments also provide strong evidence that I-PpoI contains several bound zinc ions as part of its structure.

References

Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Apr 28, 1968·Journal of Molecular Biology·B W Matthews
Apr 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·J S HanasC W Wu
Sep 1, 1995·Trends in Biochemical Sciences·A A Cooper, T H Stevens
Jan 1, 1994·Annual Review of Biochemistry·C M Joyce, T A Steitz
Jul 1, 1995·Journal of Bacteriology·M BelfortJ Z Dalgaard
Dec 1, 1993·Molecular and Cellular Biology·E L Ellison, V M Vogt
Jan 1, 1993·Annual Review of Biochemistry·A M Lambowitz, M Belfort
Sep 11, 1993·Nucleic Acids Research·S JohansenN P Willassen
Dec 22, 1995·The Journal of Biological Chemistry·M Belfort, P S Perlman
Jan 23, 1996·Biochemistry·P K Wittmayer, R T Raines
Jun 1, 1997·Nature Structural Biology·P J HeathB L Stoddard
Sep 1, 1997·Nucleic Acids Research·M Belfort, R J Roberts
Sep 1, 1940·The Biochemical Journal·D Keilin, T Mann
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

❮ Previous
Next ❯

Citations

Dec 1, 2005·Journal of Biological Physics·S N Volkov
Aug 28, 2007·Nucleic Acids Research·Jennifer L EklundRaymond J Monnat
Oct 20, 2007·Nucleic Acids Research·Jennifer H EastbergBarry L Stoddard
Dec 18, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Pilar RedondoGuillermo Montoya
Jun 11, 2014·Nature Communications·Roberto GaliziAndrea Crisanti
Nov 17, 2009·Cellular and Molecular Life Sciences : CMLS·Maria J MarcaidaGuillermo Montoya
Nov 15, 2007·The Journal of Biological Chemistry·Jesús PrietoFrancisco J Blanco

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.