Dependence of myoblast fusion on a cortical actin wall and nonmuscle myosin IIA

AbstractCell–cell fusion is a fundamental cellular process that is essential for development as well as fertilization. Myoblast fusion to form multinucleated skeletal muscle myotubes is a well studied, yet incompletely understood example of cell–cell fusion that is essential for formation of contractile skeletal muscle tissue. Studies in this report identify several novel cytoskeletal events essential to an early phase of myoblast fusion among cultured murine myoblasts. During myoblast pairing and alignment, cortical actin filaments organize into a dense actin wall structure that parallels and extends the length of the plasma membrane of the bipolar, aligned cells. As fusion progresses, gaps appear within the actin wall at sites of vesicle accumulation, the vesicles pair across the aligned myoblasts, cell–cell contacts and fusion pores form. Inhibition of nonmuscle myosin IIA (NM-MHC-IIA) motor activity prevents formation of this cortical actin wall, as well as the appearance of vesicles at a membrane proximal location, and myoblast fusion. These results suggest that early formation of a subplasmalemmal actin wall during myoblast alignment is a critical event for myoblast fusion that supports bipolar membrane alignment and temporally regulates trafficking of vesicles to the nascent fusion sites during skeletal muscle myoblast differentiation

Myosin II Light Chain Phosphorylation Regulates Membrane Localization and Apoptotic Signaling of Tumor Necrosis Factor Receptor-1

Activation of myosin II by myosin light chain kinase (MLCK) produces the force for many cellular processes including muscle contraction, mitosis, migration, and other cellular shape changes. The results of this study show that inhibition or potentiation of myosin II activation via over-expression of a dominant negative or wild type MLCK can delay or accelerate tumor necrosis factor-α (TNF)-induced apoptotic cell death in cells. Changes in the activation of caspase-8 that parallel changes in regulatory light chain phosphorylation levels reveal that myosin II motor activities regulate TNF receptor-1 (TNFR-1) signaling at an early step in the TNF death signaling pathway. Treatment of cells with either ionomycin or endotoxin (lipopolysaccharide) leads to activation of myosin II and increased translocation of TNFR-1 to the plasma membrane independent of TNF signaling. The results of these studies establish a new role for myosin II motor activity in regulating TNFR-1-mediated apoptosis through the translocation of TNFR-1 to or within the plasma membrane

A fluorescent resonant energy transfer–based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows

Approaches with high spatial and temporal resolution are required to understand the regulation of nonmuscle myosin II in vivo. Using fluorescence resonance energy transfer we have produced a novel biosensor allowing simultaneous determination of myosin light chain kinase (MLCK) localization and its [Ca2+]4/calmodulin-binding state in living cells. We observe transient recruitment of diffuse MLCK to stress fibers and its in situ activation before contraction. MLCK is highly active in the lamella of migrating cells, but not at the retracting tail. This unexpected result highlights a potential role for MLCK-mediated myosin contractility in the lamella as a driving force for migration. During cytokinesis, MLCK was enriched at the spindle equator during late metaphase, and was maximally activated just before cleavage furrow constriction. As furrow contraction was completed, active MLCK was redistributed to the poles of the daughter cells. These results show MLCK is a myosin regulator in the lamella and contractile ring, and pinpoints sites where myosin function may be mediated by other kinases

HII Region Oxygen Abundances in Starbursting Transition Dwarf Galaxies

We present empirical HII region oxygen abundances for a sample of low-luminosity starburst galaxies which are in a short lived evolutionary state. All five galaxies are characterized by centrally concentrated star formation, which is embedded in smooth stellar envelopes resembling dE-like systems. The galaxies also have small gas contents with typical M_{HI}/L_{B} ~ 0.1 resulting in gas exhaustion timescales less than 1 Gyr, even when molecular gas is considered. We find, compared to other morphologically similar systems, the galaxies of this sample have surprisingly high oxygen abundances with 12 + log(O/H) ~ 9.0. We propose that these objects are a subclass of evolved blue compact dwarfs, which have exhausted most of their gas supply while retaining their metals. We further propose that we are seeing these objects during a short phase in which they are nearing the end of their starburst activity, and could become early-type dwarfs.Comment: 13 pages, 3 figures, accepted by ApJ Letter

Anomalous Evolution of the Dwarf Galaxy HIPASS J1321-31

We present HST/WFPC2 observations of the dwarf galaxy HIPASS J1321-31. This unusual galaxy lies in the direction of the Centaurus A group of galaxies, and has a color-magnitude diagram with a distinctive red plume of luminous stars. This feature could arise from (a) a red giant branch if the galaxy were much nearer than previously recognized, (b) a peculiar asymptotic giant branch, or, (c) an ~1 Gigayear old population of intermediate mass red supergiants, which we find to be the most likely explanation. However, the lack of equally luminous blue stars requires that the star formation has dropped substantially since these stars were formed. Evidently HIPASS J1321-31 experienced an episode of enhanced star formation rather recently in its star formation history followed by a period of relative quiescence which has led to the evolution of the main sequence stars into the red supergiant branch. The stellar populations in HIPASS J1321-31 reflect a star formation history that is uncommon in star forming dwarf galaxies. This is the first time such a star formation history has been noted, although the literature contains a small number of other dwarf galaxies with similar color-magnitude diagrams. Therefore, HIPASS J1321-31 and these other galaxies represent a different path of dwarf galaxy evolution that has not been well-explored and an important probe into how dwarf galaxies evolve.Comment: 5 pages, including 3 figures and 1 table, emulateapj5/apjfonts style. Accepted by the Astrophysical Journal Letter

Evolutionary Status of Dwarf ``Transition'' Galaxies

We present deep B, R and Halpha imaging of 3 dwarf galaxies: NGC3377A, NGC4286, and IC3475. Based on previous broadband imaging and HI studies, these mixed-morphology galaxies were proposed by Sandage & Hoffman (1991) to be, respectively, a gas-rich low surface brightness Im dwarf, a nucleated dwarf that has lost most of its gas and is in transition from Im to dS0,N, and the prototypical example of a gas-poor ``huge low surface brightness'' early-type galaxy. From the combination of our broadband and Halpha imaging with the published information on the neutral gas content of these three galaxies, we find that (1) NGC3377A is a dwarf spiral; (2) NGC3377A and NGC4286 have comparable amounts of ongoing star formation, as indicated by their Halpha emission, while IC3475 has no detected HII regions to a very low limit; (3) the global star formation rates are at least a factor of 20 below that of 30 Doradus for NGC3377A and NGC4286; (4) while the amount of star formation is comparable, the distribution of star forming regions is very different between NGC3377A and NGC4286; (5) given their current star formation rates and gas contents, both NGC3377A and NGC4286 can continue to form stars for more than a Hubble time; (6) both NGC3377A and NGC4286 have integrated total B-R colors that are redder than the integrated total B-R color for IC3475, and thus it is unlikely that either galaxy will ever evolve into an IC3475 counterpart; and (7) IC3475 is too blue to be a dE. We thus conclude that we have not identified potential precursors to galaxies such as IC3475, and unless signifcant changes occur in the star formation rates, neither NGC3377A nor NGC4286 will evolve into a dwarf elliptical or dwarf spheroidal within a Hubble time.Comment: 34 pages, 6 jpg figures, 3 postscript figures, and 4 tables, uses AASTeX, ApJ, in pres

Stabilization of angiotensin-(1-7) by key substitution with a cyclic non-natural amino acid

Angiotensin-(1-7) [Ang-(1-7)], a heptapeptide hormone of the renin-angiotensin-aldosterone system (RAAS), is a promising candidate as a treatment for cancer that reflects its antiproliferative and anti-angiogenic properties. However, the peptide’s therapeutic potential is limited by the short half-life and low bioavailability resulting from rapid enzymatic metabolism by peptidases including angiotensin-converting enzyme (ACE) and dipeptidyl peptidase 3 (DPP 3). We report the facile assembly of three novel Ang-(1-7) analogues by solid-phase peptide synthesis which incorporates the cyclic non-natural δ-amino acid ACCA. The analogues containing the ACCA substitution at the site of ACE cleavage exhibit complete resistance to human ACE, while substitution at the DDP3 cleavage site provided stability against DPP 3 hydrolysis. Furthermore, the analogues retain the anti-proliferative properties of Ang-(1-7) against the 4T1 and HT-1080 cancer cell lines. These results suggest that ACCA-substituted Ang-(1-7) analogues which show resistance against proteolytic degradation by peptidases known to hydrolyze the native heptapeptide may be novel therapeutics in the treatment of cancer