Biology:Spizellomyces punctatus

From HandWiki
Short description: Chytrid fungal species

Spizellomyces punctatus
Scientific classification edit
Domain: Eukaryota
Kingdom: Fungi
Division: Chytridiomycota
Class: Chytridiomycetes
Order: Spizellomycetales
Family: Spizellomycetaceae
Genus: Spizellomyces
Species:
S. punctatus
Binomial name
Spizellomyces punctatus
(W.J. Koch) D.J.S. Barr

Spizellomyces punctatus is a chytrid fungus living in soil.[1] It is a saprotrophic fungus that colonizes decaying plant material.[2] Being an early diverging fungus, S. punctatus retains ancestral cellular features that are also found in animals and amoebae.[3] Its pathogenic relatives, Batrachochytrium dendrobatidis and B. salamandrivorans, infect amphibians and cause global biodiversity loss.[4] The pure culture of S. punctatus was first obtained by Koch (named Phlyctochytrium punctatum).[5]

Genome

The genome of S. punctatus strain DAOM BR117 was sequenced under the Origins of Multicellularity project.[6] Its genome size is about 24.13 Mb with a GC content of 47.6%. The genome has 9,424 predicted transcripts and 8,952 predicted protein-coding genes. The DDBJ/EMBL/GenBank accession number is ACOE00000000.[1]

Genetic transformation

Agrobacterium-Mediated Transformation

Genetic transformation of S. punctatus zoospores by plant pathogen Agrobacterium tumefaciens EHA105 strain is successfully established. Several selection markers have been tested. The growth of S. punctatus is not inhibited by Geneticin (G418), Puromycin, and Phleomycin D10 (Zeocin) up to 800 mg/L. 200 mg/L Hygromycin and 800 mg/L Nourseothricin (CloNAT) completely inhibit S. punctatus growth. The scientists who develop this protocol use Hygromycin as the selection marker. S. punctatus HSP70 and H2B promoters drive sufficient gene expression for Hygromycin resistance and GFP expression tested in yeast. Controlled by stronger H2B promoter, however, GFP may not be successfully folded in S. punctatus. Other fluorescent proteins, including tdTomato, mClover3, mCitrine, and mCerulean3, are functional in S. punctatus.[3]

Electroporation

A high-efficiency electroporation protocol for S.punctatus and two related chytrids species B. dendrobatidis and B. salamandrivorans has also been established. The optimal voltage for S. punctatus is 1000 V. The efficiency is about 95% using synchronized zoospores. Electroporation using unsynchronized zoospores can also reach more than 80% efficiency.[7]

Life cycle

S.punctatus globular zoospores (3–5 mm) lacks a cell wall. The zoospores can swim with a motile cilium (20–24 mm) or crawl on surfaces by actin-filled pseudopods.[3]

During encystment, the cilium is disassembled first via axoneme internalization. The initiation of this process is actin-dependent. The axoneme remains intact during internalization and the axonemal tubulin is degraded at least in part by the proteasome. The cell wall is formed after axoneme internalization. Five modes of axoneme internalization occur in S. punctatus: severing, reeling in retraction, lash-around retraction, ciliary compartment loss retraction, and vesicular retraction. First, severing is referred to as cilium detachment. Second, reeling in retraction is concurrent with or without cortical rotation and termed body-twist retraction and straight-in retraction, respectively. Third, during lash-around retraction, the cilium wraps around outside the zoospore with merging of ciliary membrane and plasma membrane. On 120 kPa fibronectin-coated hydrogels, this lash-around retraction occurs within a second. Fourth, for ciliary compartment loss retraction, ciliary membrane expansion is followed by merging of the ciliary compartment with the plasma membrane. Fifth, vesicular retraction is the creation of an axoneme loop bulge within the ciliary membrane before internalization.[8][9]

After the cilium is retracted, the cyst germinates and generates a germ tube. The germ tube is then extended to form the rhizoidal system. Finally, the cyst develops into a sporangium, a reproductive structure, and mitosis begins. After five to eight times of synchronous mitosis, 32 – 256 zoospores form in the sporangium. Ciliogenesis probably occurs before cellularization. After cellularization, the zoospores escapes from the sporangium under suitable environmental condition.[3]

The timing of the cell cycle has been quantified using the S.punctatus expressing H2B-TdTomato controlled by H2B promoter under microscopy. The retraction of the cilium and the start of encysting happen within one hour. The germ tube appears in one to three hours. The first mitosis happens in eight to twelve hours. It finishes five to eight times of synchronous mitosis in thirty hours. The average cell cycle takes about 150 minutes. Each nuclear division is completed in 1 minute.[3]

Mitochondrial 5’ tRNA editing

This species is notable for having mitochondrial 5′ tRNA editing, a rare modification that is only known to also exist in the Amoebozoa species Acanthamoeba castellanii[1] and Chytridiomycota species Harpochytrium94, Harpochytrium105, Monoblepharella15, and Hyaloraphidium curvatum.[10][11] S. punctatus mitochondrial genome encodes eight tRNAs that recognize lysine, aspartic acid, tryptophan, methionine, tyrosine, glutamine, proline, and leucine codons. tRNALeu recognizes the UAG codon as leucine instead of the stop codon.[10]

tRNAs form secondary structures that are composed of helical stems. Predicted from mtDNA, mismatches are found in the first three nucleotides of the eight tRNA acceptor stems. Sequencing of the mature mitochondrial tRNAs showed the replacement of pyrimidines or purines by purines (A to G, U to G, U to A, and C to A) that restore the base pairing. The editing sites are always restricted to the first three positions.[10][12]

The mitochondrial 5’ tRNA editing of S.punctatus has been confirmed in vitro. Using mitochondrial extract, the 5’ mismatches of synthetic tRNA transcripts are removed and nucleotides are incorporated in a 3’ to 5’ direction by using the 3’ tRNA sequence as templates. The patterns of mitochondrial 5’ tRNA editing are similar to those found in A. castellanii.[13]

Phytohormone receptor homologs

Ethylene and cytokinin receptors in plants are histidine kinases.[14] Histidine kinases in fungi are hybrid histidine kinases due to the fusion of histidine kinase/histidine kinase-like ATPase catalytic domains (HK/HATPase domains) to the receiver domain. Ethylene and cytokinin receptor homologs are also found in several flagellated and unflagellated fungal genera, including Spizellomyces. In general, these two phytohormones are signaling molecules in plant biotic interactions. Ethylene and cytokinin receptors in early diversifying fungus may play important roles in colonizing land.[2]

Opsins

Two opsin types exist:[15][16] Type 1 opsins are used by prokaryotes and by some algae (as a component of channelrhodopsins) and fungi,[17] whereas animals use type 2 opsins.[15] Type 2 opsins belong to Class A family of G-protein coupled receptors.[18] Both types are seven-transmembrane receptors and bind covalently retinal as chromophore, which turns them into photoreceptors sensing light. However, both types are not related on the sequence level.[19]

In other fungi such as Blastocladiella emersonii, a flagellated early-diverging fungus, type 1 opsins are used for phototaxis.[20] However, in S. punctatus type 1 opsins do not exist,[21] but a putative type 2 opsin. It shares with other G-protein-coupled receptors a number of conserved motifs and amino acids including the lysine corresponding to residue 296 in cattle rhodopsin,[22] which is important for retinal binding and light sensing.[23] It is as suggested by template-based structure modelling also structurally similar to animal type 2 opsins. At least computationally, it can bind retinal as chromophore. However, it prefers binding 9-cis-retinal,[22] unlike most classical animal type 2 opsins, such as cattle rhodopsin, which binds 11-cis-retinal in the dark state.[24][25][26][27][28] However, the biological function of the S. punctatus opsin is unknown.[22] Whether it is indeed a type 2 opsin is also unclear, since it is absent from a comprehensive opsin pyhlogeny that covers as many opsins as possible.[29] In principle, if it is a photoreceptor, it could have evolved light sensitivity, independently.

Fanzor endonuclease

Fanzor is a protein encoded by eukaryotic transposons and is thought to have originated from TnpB, an effector of the prokaryotic RNA-guided system known as OMEGA. TnpB is also considered the putative ancestor of Cas12, an RNA-guided endonuclease utilized in the CRISPR-Cas system. This suggests a connection between Fz, TnpB, and Cas12, despite their different roles and context in prokaryotic and eukaryotic cells. Spizellomyces punctatus was used extensively to study the structure of Fanzor.[30]

References

  1. ↑ 1.0 1.1 1.2 "Genome Sequence of Spizellomyces punctatus". Genome Announcements 4 (4). August 2016. doi:10.1128/genomeA.00849-16. PMID 27540072. 
  2. ↑ 2.0 2.1 "The Identification of Phytohormone Receptor Homologs in Early Diverging Fungi Suggests a Role for Plant Sensing in Land Colonization by Fungi". mBio 8 (1). January 2017. doi:10.1128/mBio.01739-16. PMID 28143977. 
  3. ↑ 3.0 3.1 3.2 3.3 3.4 "Genetic transformation of Spizellomyces punctatus, a resource for studying chytrid biology and evolutionary cell biology". eLife 9: e52741. May 2020. doi:10.7554/eLife.52741. PMID 32392127. 
  4. ↑ "Chytrid fungi and global amphibian declines". Nature Reviews. Microbiology 18 (6): 332–343. June 2020. doi:10.1038/s41579-020-0335-x. PMID 32099078. https://discovery.ucl.ac.uk/id/eprint/10092667/. Retrieved 2022-08-23. 
  5. ↑ "Two new chytrids in pure culture, Phlyctochytrium punctatum and Phlyctochytrium irregulare.". Journal of the Elisha Mitchell Scientific Society 73 (1): 108–122. May 1957. 
  6. ↑ "The origins of multicellularity: a multi-taxon genome initiative" (in English). Trends in Genetics 23 (3): 113–118. March 2007. doi:10.1016/j.tig.2007.01.005. PMID 17275133. 
  7. ↑ "High-efficiency electroporation of chytrid fungi". Scientific Reports 10 (1): 15145. September 2020. doi:10.1038/s41598-020-71618-2. PMID 32934254. Bibcode: 2020NatSR..1015145S. 
  8. ↑ "Studies of the Motile Cells of Chytrids. V. Flagellar Retraction in Posteriorly Uniflagellate Fungi" (in en). American Journal of Botany 55 (7): 841–859. 1968. doi:10.1002/j.1537-2197.1968.tb07442.x. ISSN 1537-2197. 
  9. ↑ "Cilium axoneme internalization and degradation in chytrid fungi". Cytoskeleton 77 (10): 365–378. October 2020. doi:10.1002/cm.21637. PMID 33103844. 
  10. ↑ 10.0 10.1 10.2 "Origin, evolution, and mechanism of 5' tRNA editing in chytridiomycete fungi". RNA 10 (8): 1191–1199. August 2004. doi:10.1261/rna.7330504. PMID 15247432. 
  11. ↑ "Fungal evolution: the case of the vanishing mitochondrion". Current Opinion in Microbiology. Host--microbe interactions: fungi / edited by Howard Bussey · Host--microbe interactions: parasites / edited by Artur Scherf · Host--microbe interactions: viruses / edited by Margaret CM Smith 8 (4): 362–369. August 2005. doi:10.1016/j.mib.2005.06.009. PMID 15993645. 
  12. ↑ "Mitochondrial tRNAs in the lower fungus Spizellomyces punctatus: tRNA editing and UAG 'stop' codons recognized as leucine". Nucleic Acids Research 25 (3): 626–632. February 1997. doi:10.1093/nar/25.3.626. PMID 9016605. 
  13. ↑ "In vitro characterization of a tRNA editing activity in the mitochondria of Spizellomyces punctatus, a Chytridiomycete fungus". The Journal of Biological Chemistry 280 (4): 2463–2470. January 2005. doi:10.1074/jbc.M411273200. PMID 15546859. 
  14. ↑ "Cytokinin and Ethylene Cell Signaling Pathways from Prokaryotes to Eukaryotes". Cells 9 (11): 2526. November 2020. doi:10.3390/cells9112526. PMID 33238457. 
  15. ↑ 15.0 15.1 Plachetzki, David C.; Fong, Caitlin R.; Oakley, Todd H. (7 July 2010). "The evolution of phototransduction from an ancestral cyclic nucleotide gated pathway". Proceedings of the Royal Society B: Biological Sciences 277 (1690): 1963–1969. doi:10.1098/rspb.2009.1797. PMID 20219739. 
  16. ↑ Fernald, R. D. (29 September 2006). "Casting a Genetic Light on the Evolution of Eyes". Science 313 (5795): 1914–1918. doi:10.1126/science.1127889. PMID 17008522. Bibcode: 2006Sci...313.1914F. 
  17. ↑ "Leptosphaeria rhodopsin: bacteriorhodopsin-like proton pump from a eukaryote". Proceedings of the National Academy of Sciences of the United States of America 102 (19): 6879–83. May 2005. doi:10.1073/pnas.0409659102. PMID 15860584. Bibcode: 2005PNAS..102.6879W. 
  18. ↑ "Structure-function of the G protein-coupled receptor superfamily". Annual Review of Pharmacology and Toxicology 53 (1): 531–556. 2013-01-06. doi:10.1146/annurev-pharmtox-032112-135923. PMID 23140243. 
  19. ↑ Findlay, JB; Pappin, DJ (15 September 1986). "The opsin family of proteins.". The Biochemical Journal 238 (3): 625–42. doi:10.1042/bj2380625. PMID 2948499. 
  20. ↑ Avelar, Gabriela Mól; Glaser, Talita; Leonard, Guy; Richards, Thomas A.; Ulrich, Henning; Gomes, Suely L. (September 2015). "A Cyclic GMP-Dependent K + Channel in the Blastocladiomycete Fungus Blastocladiella emersonii". Eukaryotic Cell 14 (9): 958–963. doi:10.1128/EC.00087-15. PMID 26150416. 
  21. ↑ Galindo, Luis Javier; Milner, David S.; Gomes, Suely Lopes; Richards, Thomas A. (July 2022). "A light-sensing system in the common ancestor of the fungi". Current Biology 32 (14): 3146–3153.e3. doi:10.1016/j.cub.2022.05.034. PMID 35675809. 
  22. ↑ 22.0 22.1 22.2 "Exploring the binding properties and structural stability of an opsin in the chytrid Spizellomyces punctatus using comparative and molecular modeling". PeerJ 5: e3206. 2017-04-27. doi:10.7717/peerj.3206. PMID 28462022. 
  23. ↑ Leung, Nicole Y.; Thakur, Dhananjay P.; Gurav, Adishthi S.; Kim, Sang Hoon; Di Pizio, Antonella; Niv, Masha Y.; Montell, Craig (April 2020). "Functions of Opsins in Drosophila Taste". Current Biology 30 (8): 1367–1379.e6. doi:10.1016/j.cub.2020.01.068. PMID 32243853. 
  24. ↑ Wald, George (July 1934). "Carotenoids and the Vitamin A Cycle in Vision". Nature 134 (3376): 65. doi:10.1038/134065a0. Bibcode: 1934Natur.134...65W. 
  25. ↑ Wald, George; Brown, Paul K.; Hubbard, Ruth; Oroshnik, William (15 July 1955). "Hindered Cis Isomers of Vitamin a and Retinene: The Structure of the Neo-B Isomer". Proceedings of the National Academy of Sciences 41 (7): 438–451. doi:10.1073/pnas.41.7.438. PMID 16589696. Bibcode: 1955PNAS...41..438W. 
  26. ↑ BROWN, PK; WALD, G (October 1956). "The neo-b isomer of vitamin A and retinene.". The Journal of Biological Chemistry 222 (2): 865–77. doi:10.1016/S0021-9258(20)89944-X. PMID 13367054. 
  27. ↑ Oroshnik, William (June 1956). "THE SYNTHESIS AND CONFIGURATION OF NEO-b VITAMIN A AND NEORETINENE b". Journal of the American Chemical Society 78 (11): 2651–2652. doi:10.1021/ja01592a095. 
  28. ↑ Oroshnik, William; Brown, Paul K.; Hubbard, Ruth; Wald, George (September 1956). "HINDERED CIS ISOMERS OF VITAMIN A AND RETINENE: THE STRUCTURE OF THE NEO-b ISOMER". Proceedings of the National Academy of Sciences 42 (9): 578–580. doi:10.1073/pnas.42.9.578. PMID 16589909. Bibcode: 1956PNAS...42..578O. 
  29. ↑ Gühmann, Martin; Porter, Megan L.; Bok, Michael J. (6 August 2022). "The Gluopsins: Opsins without the Retinal Binding Lysine". Cells 11 (15): 2441. doi:10.3390/cells11152441. PMID 35954284. 
  30. ↑ Saito, Makoto (28 June 2023). "Fanzor is a eukaryotic programmable RNA-guided endonuclease". Nature 620 (7974): 660–668. doi:10.1038/s41586-023-06356-2. PMID 37380027. Bibcode: 2023Natur.620..660S. 

Wikidata ☰ Q10676082 entry



Retrieved from "https://handwiki.org/wiki/index.php?title=Biology:Spizellomyces_punctatus&oldid=3628265"