Heme acquisition by Shu1 requires Nbr1 and proteins of the ESCRT complex in Schizosaccharomyces pombe

2019 ◽  
Vol 112 (5) ◽  
pp. 1499-1518 ◽  
Author(s):  
Thierry Mourer ◽  
Ariane Brault ◽  
Simon Labbé
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Heme Acquisition ◽  
Escrt Complex

scholarly journals FRCaMP, a Red Fluorescent Genetically Encoded Calcium Indicator Based on Calmodulin from Schizosaccharomyces Pombe Fungus

2020 ◽  
Vol 22 (1) ◽  
pp. 111
Author(s):  
Oksana M. Subach ◽  
Natalia V. Barykina ◽  
Elizaveta S. Chefanova ◽  
Anna V. Vlaskina ◽  
Vladimir P. Sotskov ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Hela Cells ◽  
Calcium Binding ◽  
Dynamic Range ◽  
Spectral Characteristics ◽  
Binding Domain ◽  
Calcium Transients ◽  
Neuronal Cultures ◽  
Decay Kinetics ◽  
Calcium Binding Domain

Red fluorescent genetically encoded calcium indicators (GECIs) have expanded the available pallet of colors used for the visualization of neuronal calcium activity in vivo. However, their calcium-binding domain is restricted by calmodulin from metazoans. In this study, we developed red GECI, called FRCaMP, using calmodulin (CaM) from Schizosaccharomyces pombe fungus as a calcium binding domain. Compared to the R-GECO1 indicator in vitro, the purified protein FRCaMP had similar spectral characteristics, brightness, and pH stability but a 1.3-fold lower ΔF/F calcium response and 2.6-fold tighter calcium affinity with Kd of 441 nM and 2.4–6.6-fold lower photostability. In the cytosol of cultured HeLa cells, FRCaMP visualized calcium transients with a ΔF/F dynamic range of 5.6, which was similar to that of R-GECO1. FRCaMP robustly visualized the spontaneous activity of neuronal cultures and had a similar ΔF/F dynamic range of 1.7 but 2.1-fold faster decay kinetics vs. NCaMP7. On electrically stimulated cultured neurons, FRCaMP demonstrated 1.8-fold faster decay kinetics and 1.7-fold lower ΔF/F values per one action potential of 0.23 compared to the NCaMP7 indicator. The fungus-originating CaM of the FRCaMP indicator version with a deleted M13-like peptide did not interact with the cytosolic environment of the HeLa cells in contrast to the metazoa-originating CaM of the similarly truncated version of the GCaMP6s indicator with a deleted M13-like peptide. Finally, we generated a split version of the FRCaMP indicator, which allowed the simultaneous detection of calcium transients and the heterodimerization of bJun/bFos interacting proteins in the nuclei of HeLa cells with a ΔF/F dynamic range of 9.4 and a contrast of 2.3–3.5, respectively.

scholarly journals Active and inactive transplacement of the M26 recombination hotspot in Schizosaccharomyces pombe.

Genetics ◽  
1995 ◽  
Vol 141 (1) ◽  
pp. 33-48
Author(s):  
J B Virgin ◽  
J Metzger ◽  
G R Smith
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Meiotic Recombination ◽  
Plasmid Dna ◽  
Context Effect ◽  
Intragenic Recombination ◽  
Multicopy Plasmid ◽  
Chromosomal Locus ◽  
Sequence Element ◽  
Recombination Hotspot ◽  
Recombination Hotspots

Abstract The ade6-M26 mutation of the fission yeast Schizosaccharomyces pombe creates a meiotic recombination hotspot that elevates ade6 intragenic recombination approximately 10-15-fold. A heptanucleotide sequence including the M26 point mutation is required but not sufficient for hotspot activity. We studied the effects of plasmid and chromosomal context on M26 hotspot activity. The M26 hotspot was inactive on a multicopy plasmid containing M26 embedded within 3.0 or 5.9 kb of ade6 DNA. Random S. pombe genomic fragments totaling approximately 7 Mb did not activate the M26 hotspot on a plasmid. M26 hotspot activity was maintained when 3.0-, 4.4-, and 5.9-kb ade6-M26 DNA fragments, with various amounts of non-S. pombe plasmid DNA, were integrated at the ura4 chromosomal locus, but only in certain configurations relative to the ura4 gene and the cointegrated plasmid DNA. Several integrations created new M26-independent recombination hotspots. In all cases the non-ade6 DNA was located > 1 kb from the M26 site, and in some cases > 2 kb. Because the chromosomal context effect was transmitted over large distances, and did not appear to be mediated by a single discrete DNA sequence element, we infer that the local chromatin structure has a pronounced effect on M26 hotspot activity.

scholarly journals Identification of a human homologue of the Schizosaccharomyces pombe rad17 checkpoint gene.

1999 ◽  
Vol 274 (34) ◽  
pp. 24438-24439
Author(s):  
Andrew E. Parker ◽  
Inez Van de Weyer ◽  
Marc C. Laus ◽  
Peter Verhasselt ◽  
Walter H.M.L. Luyten
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Human Homologue ◽  
Checkpoint Gene

scholarly journals Partial purification of RNase P from Schizosaccharomyces pombe.

1981 ◽  
Vol 256 (10) ◽  
pp. 5058-5063
Author(s):  
L. Kline ◽  
S. Nishikawa ◽  
D. Söll
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Rnase P ◽  
Partial Purification
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