Essential nucleotide- and protein-dependent functions of Actb/ß-actin.

The highly similar cytoplasmic ß- and γ-actins differ by only four functionally similar amino acids, yet previous in vitro and in vivo data suggest that they support unique functions due to striking phenotypic differences between Actb and Actg1 null mouse and cell models. To determine whether the four amino acid variances were responsible for the functional differences between cytoplasmic actins, we gene edited the endogenous mouse Actb locus to translate γ-actin protein. The resulting mice and primary embryonic fibroblasts completely lacked ß-actin protein, but were viable and did not present with the most overt and severe cell and organismal phenotypes observed with gene knockout. Nonetheless, the edited mice exhibited progressive high-frequency hearing loss and degeneration of actin-based stereocilia as previously reported for hair cell-specific Actb knockout mice. Thus, ß-actin protein is not required for general cellular functions, but is necessary to maintain auditory stereocilia.

Stereocilia morphogenesis and maintenance through regulation of actin stability.

Stereocilia are actin-based protrusions on auditory and vestibular sensory cells that are required for hearing and balance. They convert physical force from sound, head movement or gravity into an electrical signal, a process that is called mechanoelectrical transduction. This function depends on the ability of sensory cells to grow stereocilia of defined lengths. These protrusions form a bundle with a highly precise geometry that is required to detect nanoscale movements encountered in the inner ear. Congenital or progressive stereocilia degeneration causes hearing loss. Thus, understanding stereocilia hair bundle structure, development, and maintenance is pivotal to understanding the pathogenesis of deafness. Stereocilia cores are made from a tightly packed array of parallel, crosslinked actin filaments, the length and stability of which are regulated in part by myosin motors, actin crosslinkers and capping proteins. This review aims to describe stereocilia actin regulation in the context of an emerging "tip turnover" model where actin assembles and disassembles at stereocilia tips while the remainder of the core is exceptionally stable.

A naphthalene-based heterobimetallic triazolylidene IrIII/PdII complex: regioselective to regiospecific C-H activation, tandem catalysis and a copper-free Sonogashira reaction.

Heterobimetallic complexes featuring mesoionic carbene (MIC) donor ligands are gaining enormous popularity in tandem catalysis owing to the combined action of two different metal centers during catalysis. A rare version of the heterobimetallic PdII/IrIII complex possessing a cyclometalated mesoionic carbene (MIC) ligand is presented along with the analogous homodinuclear PdII complex. A sterically controlled regiospecific cyclometalation towards the formation of a six-membered ring complex over a five-membered ring complex has been performed using a naphthalene-based bis-MIC ligand platform. The interplay between regioselective vs. regiospecific C-H bond activation for the synthesis of cyclometalated IrIII complexes has also been demonstrated using the corresponding naphthyl-derived mono-imidazolylidene ligand. Both homodinuclear PdII and heterobimetallic PdII/IrIII complexes have been characterized using standard spectroscopic techniques including 1H, 13C{1H}, 2D correlation NMR spectroscopy and ESI mass spectrometry. The structure of the cyclometalated heterobimetallic complex has been established by single crystal XRD. The heterobimetallic complex has been employed as a pre-catalyst in the tandem Suzuki-Miyaura/transfer hydrogenation reaction and the homobimetallic PdII complex has been successfully employed as a catalyst in both the Sonogashira coupling and α-arylation of 1-methyl-2-oxindole.

Strategic Approach Towards Plastic Waste Valorization: Challenges and Promising Chemical Upcycling Possibilities.

Plastic waste, which is one of the major sources of pollution in the landfills and oceans, has raised global concern, primarily due to the huge production rate, high durability, and the lack of utilization of the available waste management techniques. Recycling methods are preferable to reduce the impact of plastic pollution to some extent. However, most of the recycling techniques are associated with different drawbacks, high cost and downgrading of product quality being among the notable ones. The sustainable option here is to upcycle the plastic waste to create high-value materials to compensate for the cost of production. Several upcycling techniques are constantly being investigated and explored, which is currently the only economical option to resolve the plastic waste issue. This Review provides a comprehensive insight on the promising chemical routes available for upcycling of the most widely used plastic and mixed plastic wastes. The challenges inherent to these processes, the recent advances, and the significant role of the science and research community in resolving these issues are further emphasized.

Actin at stereocilia tips is regulated by mechanotransduction and ADF/cofilin.

Stereocilia on auditory sensory cells are actin-based protrusions that mechanotransduce sound into an electrical signal. These stereocilia are arranged into a bundle with three rows of increasing length to form a staircase-like morphology that is required for hearing. Stereocilia in the shorter rows, but not the tallest row, are mechanotransducing because they have force-sensitive channels localized at their tips. The onset of mechanotransduction during mouse postnatal development refines stereocilia length and width. However, it is unclear how actin is differentially regulated between stereocilia in the tallest row of the bundle and the shorter, mechanotransducing rows. Here, we show actin turnover is increased at the tips of mechanotransducing stereocilia during bundle maturation. Correspondingly, from birth to postnatal day 6, these stereocilia had increasing amounts of available actin barbed ends, where monomers can be added or lost readily, as compared with the non-mechanotransducing stereocilia in the tallest row. The increase in available barbed ends depended on both mechanotransduction and MYO15 or EPS8, which are required for the normal specification and elongation of the tallest row of stereocilia. We also found that loss of the F-actin-severing proteins ADF and cofilin-1 decreased barbed end availability at stereocilia tips. These proteins enriched at mechanotransducing stereocilia tips, and their localization was perturbed by the loss of mechanotransduction, MYO15, or EPS8. Finally, stereocilia lengths and widths were dysregulated in Adf and Cfl1 mutants. Together, these data show that actin is remodeled, likely by a severing mechanism, in response to mechanotransduction.

The stable actin core of mechanosensory stereocilia features continuous turnover of actin cross-linkers.

Stereocilia are mechanosensitive protrusions on the surfaces of sensory hair cells in the inner ear that detect sound, gravity, and head movement. Their cores are composed of parallel actin filaments that are cross-linked and stabilized by several actin-binding proteins, including fascin-2, plastin-1, espin, and XIRP2. The actin filaments are the most stable known, with actin turnover primarily occurring at the stereocilia tips. While stereocilia actin dynamics has been well studied, little is known about the behavior of the actin cross-linking proteins, which are the most abundant type of protein in stereocilia after actin and are critical for stereocilia morphogenesis and maintenance. Here, we developed a novel transgenic mouse to monitor EGFP-fascin-2 incorporation . In contrast to actin, EGFP-fascin-2 readily enters the stereocilia core. We also compared the effect of EGFP-fascin-2 expression on developing and mature stereocilia. When it was induced during hair cell development, we observed increases in both stereocilia length and width. Interestingly, stereocilia size was not affected when EGFP-fascin-2 was induced in adult stereocilia. Regardless of the time of induction, EGFP-fascin-2 displaced both espin and plastin-1 from stereocilia. Altering the actin cross-linker composition, even as the actin filaments exhibit little to no turnover, provides a mechanism for ongoing remodeling and repair important for stereocilia homeostasis.