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1. Spectroscopic study of the 4f ⁷ 6s ^{2 8} S 7/2∘ – 4f ⁷ ( ⁸ S ^° ) 6s6p( ¹ P ^° ) ⁸ P _5/2,7/2 transitions in neutral europium-151 and europium-153: absolute frequency and hyperfine structure

   https://doi.org/10.1364/JOSAB.521181  

   Maruko, C.; Çölmek, N_N; Herd, M_T; Ahrendsen, K_J; Cabrales, B.; Cannon, G.; Davis, E.; Guo, X_Y; Karani, T.; Wallace, A.; et al (April 2024, Journal of the Optical Society of America B)

   We report on spectroscopic measurements on the 4f⁷6s²⁸S_7/2^∘−4f⁷(⁸S^∘)6s6p(¹P^∘)⁸P_5/2,7/2transitions at 466.32 nm and 462.85 nm, respectively, in neutral europium-151 and europium-153. The center of gravity frequencies for the 151 and 153 isotopes for both transitions are reported for the first time using saturated absorption spectroscopy. For the 6s6p(¹P^∘)⁸P_5/2state, the center of gravity frequencies were found to be 642,894,493.3(4) MHz and 642,891,693.3(9) MHz for the 151 and 153 isotopes, respectively. The hyperfine constants for the upper state were found to beA(151)=−157.01(3)MHz,B(151)=74.5(4)MHz andA(153)=−69.43(14)MHz,B(153)=191.0(26)MHz. These hyperfine values are all consistent with previously published results except forB(151) that has a small discrepancy. The isotope shift was found to be 2799.54(20) MHz, a small discrepancy with previously published results. For the 6s6p(¹P^∘)⁸P_7/2state, the center of gravity frequencies were found to be 647,708,930.6(6) MHz and 647,705,958.4(26) MHz for the 151 and 153 isotopes, respectively. The hyperfine constants for the upper state were found to beA(151)=−218.66(4)MHz,B(151)=−293.4(8)MHz andA(153)=−97.15(13)MHz,B(153)=−750(3)MHz. These values are all consistent with previously published results except forA(151) that has a small discrepancy. The isotope shift was found to be 2972.8(5) MHz, a small discrepancy with previously measured results. 

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2. Alternate routes to mnm ⁵ s ² U synthesis in Gram-positive bacteria

   https://doi.org/10.1128/jb.00452-23  

   Jaroch, Marshall; Sun, Guangxin; Tsui, Ho-Ching Tiffany; Reed, Colbie; Sun, Jingjing; Jörg, Marko; Winkler, Malcolm E; Rice, Kelly C; Dziergowska, Agnieszka; Stich, Troy A; et al (April 2024, Journal of Bacteriology)

   Henkin, Tina M (Ed.)

   The wobble bases of tRNAs that decode split codons are often heavily modified. In bacteria, tRNA^{Glu, Gln, Asp}contains a variety of xnm⁵s²U derivatives. The synthesis pathway for these modifications is complex and fully elucidated only in a handful of organisms, including the Gram-negativeEscherichia coliK12 model. Despite the ubiquitous presence of mnm⁵s²U modification, genomic analysis shows the absence ofmnmCorthologous genes, suggesting the occurrence of alternate biosynthetic schemes for the conversion of cmnm⁵s²U to mnm⁵s²U. Using a combination of comparative genomics and genetic studies, a member of the YtqA subgroup of the radical Sam superfamily was found to be involved in the synthesis of mnm⁵s²U in bothBacillus subtilisandStreptococcus mutans. This protein, renamed MnmL, is encoded in an operon with the recently discovered MnmM methylase involved in the methylation of the pathway intermediate nm⁵s²U into mnm⁵s²U inB. subtilis. Analysis of tRNA modifications of bothS. mutansandStreptococcus pneumoniaeshows that growth conditions and genetic backgrounds influence the ratios of pathway intermediates owing to regulatory loops that are not yet understood. The MnmLM pathway is widespread along the bacterial tree, with some phyla, such as Bacilli, relying exclusively on these two enzymes. Although mechanistic details of these newly discovered components are not fully resolved, the occurrence of fusion proteins, alternate arrangements of biosynthetic components, and loss of biosynthetic branches provide examples of biosynthetic diversity to retain a conserved tRNA modification in Nature.IMPORTANCEThe xnm⁵s²U modifications found in several tRNAs at the wobble base position are widespread in bacteria where they have an important role in decoding efficiency and accuracy. This work identifies a novel enzyme (MnmL) that is a member of a subgroup of the very versatile radical SAM superfamily and is involved in the synthesis of mnm⁵s²U in several Gram-positive bacteria, including human pathogens. This is another novel example of a non-orthologous displacement in the field of tRNA modification synthesis, showing how different solutions evolve to retain U34 tRNA modifications. 

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3. Formation of Stilbene Azo‐Dimer by Direct Irradiation of p‐Azidostilbene ^†

   https://doi.org/10.1111/php.13659  

   Osisioma, Onyinye; Patton, Leanna J.; Merugu, Rajkumar; Govorov, Dmitrii; Milbrandt, Margaret A.; Jarus, Cassandra; Karney, William L.; Gudmundsdottir, Anna D. (July 2022, Photochemistry and Photobiology)

   ABSTRACT Triplet arylnitrenes may provide direct access to aryl azo‐dimers, which have broad commercial applicability. Herein, the photolysis ofp‐azidostilbene (1) in argon‐saturated methanol yielded stilbene azo‐dimer (2) through the dimerization of tripletp‐nitrenostilbene (³1N). The formation of³1Nwas verified by electron paramagnetic resonance spectroscopy and absorption spectroscopy (λ_max ~ 375 nm) in cryogenic 2‐methyltetrahydrofuran matrices. At ambient temperature, laser flash photolysis of1in methanol formed³1N(λ_max ~ 370 nm, 2.85 × 10⁷ s⁻¹). On shorter timescales, a transient absorption (λ_max ~ 390 nm) that decayed with a similar rate constant (3.11 × 10⁷ s⁻¹) was assigned to a triplet excited state (T) of1. Density functional theory calculations yielded three configurations for T of1, with the unpaired electrons on the azido (T_A) or stilbene moiety (T_Tw, twisted and T_Fl, flat). The transient was assigned to T_Twbased on its calculated spectrum. CASPT2 calculations gave a singlet–triplet energy gap of 16.6 kcal mol⁻¹for1 N; thus, intersystem crossing of¹1Nto³1Nis unlikely at ambient temperature, supporting the formation of³1Nfrom T of1. Thus, sustainable synthetic methods for aryl azo‐dimers can be developed using the visible‐light irradiation of aryl azides to form triplet arylnitrenes. 

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4. A Nonheme Mononuclear {FeNO} ⁷ Complex that Produces N ₂ O in the Absence of an Exogenous Reductant

   https://doi.org/10.1002/anie.202109062  

   Dey, Aniruddha; Gordon, Jesse_B; Albert, Therese; Sabuncu, Sinan; Siegler, Maxime_A; MacMillan, Samantha_N; Lancaster, Kyle_M; Moënne‐Loccoz, Pierre; Goldberg, David_P (August 2021, Angewandte Chemie International Edition)

   Abstract A new nonheme iron(II) complex, Fe^II(Me₃TACN)((OSi^Ph2)₂O) (1), is reported. Reaction of1with NO_(g)gives a stable mononitrosyl complex Fe(NO)(Me₃TACN)((OSi^Ph2)₂O) (2), which was characterized by Mössbauer (δ=0.52 mm s⁻¹, |ΔE_Q|=0.80 mm s⁻¹), EPR (S=3/2), resonance Raman (RR) and Fe K‐edge X‐ray absorption spectroscopies. The data show that2is an {FeNO}⁷complex with anS=3/2 spin ground state. The RR spectrum (λ_exc=458 nm) of2combined with isotopic labeling (¹⁵N,¹⁸O) reveals ν(N‐O)=1680 cm⁻¹, which is highly activated, and is a nearly identical match to that seen for the reactive mononitrosyl intermediate in the nonheme iron enzyme FDPnor (ν(NO)=1681 cm⁻¹). Complex2reacts rapidly with H₂O in THF to produce the N‐N coupled product N₂O, providing the first example of a mononuclear nonheme iron complex that is capable of converting NO to N₂O in the absence of an exogenous reductant. 

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5. Phosphatidylinositol 3, 5‐bisphosphate regulates Ca ²⁺ transport during yeast vacuolar fusion through the Ca ²⁺ ATPase Pmc1

   https://doi.org/10.1111/tra.12736  

   Miner, Gregory E.; Sullivan, Katherine D.; Zhang, Chi; Rivera‐Kohr, David; Guo, Annie; Hurst, Logan R.; Ellis, Ez C.; Starr, Matthew L.; Jones, Brandon C.; Fratti, Rutilio A. (June 2020, Traffic)

   Abstract The transport of Ca²⁺across membranes precedes the fusion and fission of various lipid bilayers. Yeast vacuoles under hyperosmotic stress become fragmented through fission events that requires the release of Ca²⁺stores through the TRP channel Yvc1. This requires the phosphorylation of phosphatidylinositol‐3‐phosphate (PI3P) by the PI3P‐5‐kinase Fab1 to produce transient PI(3,5)P₂pools. Ca²⁺is also released during vacuole fusion upontrans‐SNARE complex assembly, however, its role remains unclear. The effect of PI(3,5)P₂on Ca²⁺flux during fusion was independent of Yvc1. Here, we show that while low levels of PI(3,5)P₂were required for Ca²⁺uptake into the vacuole, increased concentrations abolished Ca²⁺efflux. This was as shown by the addition of exogenous dioctanoyl PI(3,5)P₂or increased endogenous production of by the hyperactivefab1^T2250Amutant. In contrast, the lack of PI(3,5)P₂on vacuoles from the kinase deadfab1^EEEmutant showed delayed and decreased Ca²⁺uptake. The effects of PI(3,5)P₂were linked to the Ca²⁺pump Pmc1, as its deletion rendered vacuoles resistant to the effects of excess PI(3,5)P₂. Experiments with Verapamil inhibited Ca²⁺uptake when added at the start of the assay, while adding it after Ca²⁺had been taken up resulted in the rapid expulsion of Ca²⁺. Vacuoles lacking both Pmc1 and the H⁺/Ca²⁺exchanger Vcx1 lacked the ability to take up Ca²⁺and instead expelled it upon the addition of ATP. Together these data suggest that a balance of efflux and uptake compete during the fusion pathway and that the levels of PI(3,5)P₂can modulate which path predominates. 

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