Alam ST et al. (2021) J Photochem Photobiol B "Natural photosensitizers from Tripterygium wilfordii and their antimicrobial photodynamic therapeutic effects in a Caenorhabditis elegans model."

Kwon HC, Son JD, Hwang H, Park JS, Kwon J, Kang K, Alam ST, Nguyen UTT

[J Photochem Photobiol B, 2021]

Tripterygium wilfordii Hook. f. is a traditional medicinal plant and has long been used in East Asia to treat many diseases. However, the extract and active components have never been investigated as potential photosensitizers for photodynamic treatment to kill pathogenic microorganisms. Here, the antimicrobial photodynamic treatment (APDT) effects of the extract, fractions, and compounds of T. wilfordii were evaluated in vitro and in vivo. Ethanolic extract (TWE) and the photosensitizer-enriched fraction (TW-F5) were prepared from dried T. wilfordii. Six active compounds were isolated from TW-F5 by semipreparative high-performance liquid chromatography, and their chemical structures were characterized through spectroscopic and spectrometric analysis. The singlet oxygen from extracts, fractions, and compounds was measured by using the imidazole-N,N-dimethyl-4-nitrosoaniline method. These extracts, fractions, and compounds were used as photosensitizers for the inactivation of bacteria and fungi by red light at 660nm. The in vitro APDT effects were also evaluated in the model animal Caenorhabditis elegans. APDT with TWE showed effective antimicrobial activity against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Candida albicans. TW-F5, consisting of six pheophorbide compounds, also showed strong APDT activity. The photosensitizers were taken up into the bacterial cells and induced intracellular ROS production by APDT. TWE and TW-F5 also induced a strong APDT effect in vitro against skin pathogens, including Staphylococcus epidermidis and Streptococcus pyogenes. We evaluated the APDT effects of TWE and TW-F5 in C. elegans infected with various pathogens and found that PDT effectively controlled pathogenic bacteria without strong side effects. APDT reversed the growth retardation of worms induced by pathogen infection and decreased the viable pathogenic bacterial numbers associated with C. elegans. Finally, APDT with TWE increased the survivability of C. elegans infected with S. pyogenes. In summary, TWE and TW-F5 were found to be effective antimicrobial photosensitizers in PDT.

NH3027

Caenorhabditis elegans

Chen JS et al. (1997) J Mol Evol "Extensive sequence conservation among insect, nematode, and vertebrate vitellogenins reveals ancient common ancestry."

Chen JS, Raikhel AS, Sappington TW

[J Mol Evol, 1997]

The eggs of most oviparous animals are provisioned with a class of protein called vitellogenin (Vg) which is stored as the major component of yolk. Until recently, deduced amino acid sequences were available only from vertebrate and nematode Vgs, which proved to be homologous. The sequences of several insect Vgs are now known, but early attempts at pairwise alignments with vertebrate and nematode Vgs have been problematic, leading to conflicting conclusions about how closely insect Vgs are related to the others. In this paper we demonstrate that insect VE sequences can be confidently aligned with one another along their entire lengths and with multiple vertebrate and nematode Vg sequences along most of their spans. Although divergence is high, conservation among insect, vertebrate, and nematode Vg sequences is widespread with a preponderance of glycine, proline, and cysteine residues among strictly conserved amino acids, establishing conclusively that Vgs from the three phyla are homologous. Areas of least-certain alignment are primarily in and around insect and vertebrate polyserine domains which are not homologous. Phylogenetic reconstructions of Vgs based on sequence identities indicate that the insect lineage is the most diverged and that the mammalian serum protein, apolipoprotein B-100, arose from a Vg ancestor after the nematode/vertebrate divergence.

protein: tw-PA

Drosophila melanogaster

Flybase gene name is tw-PA

TY5640

Pristionchus pacificus

tw

Drosophila melanogaster

twisted (tw) encodes a protein that transfers a mannose to the Ser/Thr residues of dystroglycan, via forming an heterodimer with the product of rt. Its roles include myogenesis, muscle architecture and cell adhesion.

rt

Drosophila melanogaster

rotated abdomen (rt) encodes a protein that transfers a mannose to the Ser/Thr residues of the product of Dg, via forming an heterodimer with the product of tw. Its roles include myogenesis, muscle architecture and cell adhesion.

Terskikh A et al. (2000) Science ""Fluorescent timer": protein that changes color with time."

Matz M, Ermakova G, Siebert P, Kim SK, Lukyanov S, Kajava AV, Weissman I, Zaraisky A, Terskikh A, Tan PBO, Fradkov A, Zhao X

[Science, 2000]

We generated a mutant of the red fluorescent protein drFP583. The mutant (E5) changes its fluorescence from green to red over time. The rate of color conversion is independent of protein concentration and therefore can be used to trace time-dependent expression. We used in vivo labeling with E5 to measure expression from the heat shock-dependent promoter in Caenorhabditis elegans and from the Otx-2 promoter in developing Xenopus embryos. Thus, E5 is a "fluorescent timer" that can be used to monitor both activation and down-regulation of target promoters on the whole-organism scale.AD - School of Medicine, Stanford University, Stanford, CA 94305, USA. Alexey.Terskikh@Stanford.eduFAU - Terskikh, AAU - Terskikh AFAU - Fradkov, AAU - Fradkov AFAU - Ermakova, GAU - Ermakova GFAU - Zaraisky, AAU - Zaraisky AFAU - Tan, PAU - Tan PFAU - Kajava, A VAU - Kajava AVFAU - Zhao, XAU - Zhao XFAU - Lukyanov, SAU - Lukyanov SFAU - Matz, MAU - Matz MFAU - Kim, SAU - Kim SFAU - Weissman, IAU - Weissman IFAU - Siebert, PAU - Siebert PLA - engID - 1 RO3 TW01362-01/TW/FICPT - Journal ArticleCY - UNITED STATESTA - ScienceJID - 0404511RN - 0 (Heat-Shock Proteins)RN - 0 (Luminescent Proteins)RN - 0 (Nerve Tissue Proteins)RN - 0 (Otx2 protein)RN - 0 (Trans-Activators)RN - 0 (red fluorescent protein)SB - IM

Heestand, Bree et al. (2021) International Worm Meeting "Adult longevity of late-generation Piwi/prg-1 mutants"

Lister-Shimauchi, Evan, Heestand, Bree, Ahmed, Shawn, Kennedy, Sophia, Simon, Matt

[International Worm Meeting, 2021]

The C. elegans Piwi Argonaute protein PRG-1 associated with piRNAs to preventing germ cells from expressing foreign genetic elements such as transposons. PRG-1 promotes germ cell immortality and lines of prg-1 mutants are initially fertile but become sterile if they are grown for many generations. prg-1 mutant sterility is a form of reproductive arrest (1), which may resemble reproductive arrest in response to environmental stresses such as starvation (2). We found that early-generation prg-1 mutants have normal lifespans and fertility, while later generations had reduced fertility yet longer lifespans. Mutation of the stress response transcription factor Daf-16/FOXO, caused the longevity phenotype of late-generation prg-1 mutants to disappear, leading to the conclusion that prg-1 mutant longevity may be a hormetic stress response. Phenotypes that arise from epigenetic defects can be inherited for at least 3 generations (3,4), and we found that longevity of late-generation prg-1 mutants is inherited by F1 but not F2 cross progeny. We are currently asking if the daf-12 germline signaling pathway that regulates aging is required of prg-1 mutant longevity. Overall, our results imply that the longevity of late-generation prg-1 mutant is distinct from the longevity that is transmitted for several generations by germ cells of mutants deficient for the ASH-2/SET-2/trithorax transcriptional activation complex (3,4). 1. Heestand, B., Simon, M., Frenk, S., Titov, D. & Ahmed, S. Transgenerational Sterility of Piwi Mutants Represents a Dynamic Form of Adult Reproductive Diapause. Cell Rep. 23, 156-171 (2018). 2. Angelo, G. & Van Gilst, M. R. Starvation Protects Germline Stem Cells and Extends Reproductive Longevity in C. elegans. Science 326, 954-958 (2009). 3. Greer, E. L. et al. Transgenerational epigenetic inheritance of longevity in Caenorhabditis elegans. Nature 479, 365-371 (2011). 4. Lee TW, David HS, Engstrom AK, Carpenter BS, Katz DJ. Repressive H3K9me2 protects lifespan against the transgenerational burden of COMPASS activity in C. elegans. Elife (2019). doi: 10.7554/eLife.48498

Wang, Jennifer T. et al. (2013) International Worm Meeting "Asymmetric segregation of P granules requires granule remodeling by two novel serine-rich proteins."

Seydoux, Geraldine, Wang, Jennifer T.

[International Worm Meeting, 2013]

P granules are RNA-rich granules that are found exclusively in the germline. In adult germ cells, P granules are mostly perinuclear and stable. In contrast, in early embryos, P granules are cytoplasmic and highly dynamic. Activation of P granule dynamics in embryos is required to segregate P granules asymmetrically to the nascent germline [1,2]. We have found that activation of P granule dynamics requires P granule remodeling by two serine-rich proteins. We used a GFP::PGL-1 fusion to analyze P granule dynamics during the oocyte-to-embryo transition. Oocyte P granules are disassembled during ovulation and reappear during meiosis throughout the cytoplasm of the newly fertilized zygote. A similar disassembly/reassembly cycle is repeated in the zygote at each mitotic division, except that reassembly occurs preferentially in the cytoplasm destined for the germline blastomere. We have found that depletion of GEI-12 and its paralog C36C9.1 blocks the disassembly/reassembly cycle. In gei-12+C36C9.1(RNAi), oocyte P granules persist through ovulation and the first mitotic division, and do not segregate asymmetrically. Formation of new P granules is also blocked. GEI-12 and C36C9.1 are 70% identical serine-rich proteins with no recognizable motifs. Sequence analyses indicate that these proteins have low-complexity sequence (LCS) throughout their length. LCS domains have recently been proposed to form dynamic fibers that hold RNA granules together [3,4]. Consistent with functioning as a P granule scaffold, GEI-12 associates with P granules after reassembly in embryos. Deconvolution confocal microscopy suggests that, in the reassembled P granules, GEI-12 localizes to a core that joins together several small PGL-1 granules. We conclude that 1) GEI-12 and C36C9.1 remodel P granules during the oocyte-to-embryo transition and 2) P granule remodeling is essential for the asymmetric segregation of P granules in embryos. 1. Brangwynne CP et al (2009). Science 324, 1729 2. Gallo C et al (2010). Science 330(6011) 1685 3. Kato M et al (2012). Cell 149(4), 753-767 4. Han TW et al (2012). Cell 149(4), 768-779.