MT1-MMP (MMP-14) is a multifunctional protease that regulates ECM degradation, activation of other proteases, and a variety of cellular processes, including migration and viability in physiological and pathological contexts. Both the localization and signal transduction capabilities of MT1-MMP are dependent on its cytoplasmic domain that constitutes the final 20 C-terminal amino acids, while the rest of the protease is extracellular. In this review, we summarize the ways in which the cytoplasmic tail is involved in regulating and enacting the functions of MT1-MMP. We also provide an overview of known interactors of the MT1-MMP cytoplasmic tail and the functional significance of these interactions, as well as further insight into the mechanisms of cellular adhesion and invasion that are regulated by the cytoplasmic tail.

• Klíčová slova
• MT1-MMP, cell invasion, intracellular trafficking, matrix metalloproteinases, post-translational modifications,
• MeSH
• buněčná adheze MeSH
• matrixová metaloproteinasa 14 * metabolismus   MeSH
• pohyb buněk MeSH
• signální transdukce * MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• matrixová metaloproteinasa 14 * MeSH

The vanilloid receptor [transient receptor potential (TRP)V1, also known as VR1] is a member of the TRP channel family. These receptors share a significant sequence homology, a similar predicted structure with six transmembrane-spanning domains (S1-S6), a pore-forming region between S5 and S6, and the cytoplasmically oriented C- and N-terminal regions. Although structural/functional studies have identified some of the key amino acids influencing the gating of the TRPV1 ion channel, the possible contributions of terminal regions to vanilloid receptor function remain elusive. In the present study, C-terminal truncations of rat TRPV1 have been constructed to characterize the contribution of the cytoplasmic C-terminal region to TRPV1 function and to delineate the minimum amount of C tail necessary to form a functional channel. The truncation of 31 residues was sufficient to induce changes in functional properties of TRPV1 channel. More pronounced effects of C-terminal truncation were seen in mutants lacking the final 72 aa. These changes were characterized by a decline of capsaicin-, pH-, and heat-sensitivity; progressive reduction of the activation thermal threshold (from 41.5 to 28.6 degrees C); and slowing of the activation rate of heat-evoked membrane currents (Q10 from 25.6 to 4.7). The voltage-induced currents of the truncated mutants exhibited a slower onset, markedly reduced outward rectification, and significantly smaller peak tail current amplitudes. Truncation of the entire TRPV1 C-terminal domain (155 residues) resulted in a nonfunctional channel. These results indicate that the cytoplasmic COOH-terminal domain strongly influences the TRPV1 channel activity, and that the distal half of this structural domain confers specific thermal sensitivity.

• MeSH
• buněčné linie MeSH
• elektrická vodivost MeSH
• imunohistochemie MeSH
• kapsaicin farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• metoda terčíkového zámku MeSH
• molekulární modely MeSH
• molekulární sekvence - údaje MeSH
• mutace MeSH
• proteinkinasa C metabolismus   MeSH
• protony MeSH
• receptory léků chemie   genetika   fyziologie   MeSH
• sekvence aminokyselin MeSH
• sekvenční delece MeSH
• sekvenční seřazení MeSH
• serin genetika   MeSH
• terciární struktura proteinů MeSH
• vysoká teplota MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kapsaicin MeSH
• proteinkinasa C MeSH
• protony MeSH
• receptory léků MeSH
• serin MeSH

OBJECTIVE: Autosomal dominant hypocalcemia (ADH) is a rare disorder caused by activating mutations of the calcium-sensing receptor (CASR). The treatment of ADH patients with 1α-hydroxylated vitamin D derivatives can cause hypercalciuria leading to nephrocalcinosis. DESIGN AND METHODS: We studied a girl who presented with hypoparathyroidism and asymptomatic hypocalcemia at age 2.5 years. Mutations of CASR were investigated by DNA sequencing. Functional analyses of mutant and WT CASRs were done in transiently transfected human embryonic kidney (HEK293) cells. RESULTS: The proband and her father are heterozygous for an eight-nucleotide deletion c.2703_2710delCCTTGGAG in the CASR encoding the intracellular domain of the protein. Transient expression of CASR constructs in kidney cells in vitro suggested greater cell surface expression of the mutant receptor with a left-shifted extracellular calcium dose-response curve relative to that of the WT receptor consistent with gain of function. Initial treatment of the patient with calcitriol led to increased urinary calcium excretion. Evaluation for mosaicism in the paternal grandparents of the proband was negative. CONCLUSIONS: We describe a novel naturally occurring deletion mutation within the CASR that apparently arose de novo in the father of the ADH proband. Functional analysis suggests that the cytoplasmic tail of the CASR contains determinants that regulate the attenuation of signal transduction. Early molecular analysis of the CASR gene in patients with isolated idiopathic hypoparathyroidism is recommended because of its relevance to clinical outcome and treatment choice. In ADH patients, calcium supplementation and low-dose cholecalciferol avoids hypocalcemic symptoms without compromising renal function.

• MeSH
• cytoplazma MeSH
• dominantní geny * MeSH
• dospělí MeSH
• HEK293 buňky MeSH
• heterozygot MeSH
• hyperkalciurie genetika   patologie   MeSH
• hypokalcemie genetika   patologie   MeSH
• hypoparatyreóza vrozené   genetika   patologie   MeSH
• lidé MeSH
• nesmyslný kodon genetika   MeSH
• předškolní dítě MeSH
• receptory "calcium-sensing" chemie   genetika   MeSH
• rodina MeSH
• rodokmen MeSH
• sekvence nukleotidů MeSH
• sekvenční delece * MeSH
• terciární struktura proteinů genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nesmyslný kodon MeSH
• receptory "calcium-sensing" MeSH

Osteoblasts orchestrate bone formation through the secretion of type I collagen and other constituents of the matrix on which hydroxyapatite crystals mineralize. Here, we show that TENT5A, whose mutations were found in congenital bone disease osteogenesis imperfecta patients, is a cytoplasmic poly(A) polymerase playing a crucial role in regulating bone mineralization. Direct RNA sequencing revealed that TENT5A is induced during osteoblast differentiation and polyadenylates mRNAs encoding Col1α1, Col1α2, and other secreted proteins involved in osteogenesis, increasing their expression. We postulate that TENT5A, possibly together with its paralog TENT5C, is responsible for the wave of cytoplasmic polyadenylation of mRNAs encoding secreted proteins occurring during bone mineralization. Importantly, the Tent5a knockout (KO) mouse line displays bone fragility and skeletal hypomineralization phenotype resulting from quantitative and qualitative collagen defects. Thus, we report a biologically relevant posttranscriptional regulator of collagen production and, more generally, bone formation.

• Klíčová slova
• FAM46A, Nanopore, TENT5A, collagen, direct RNA sequencing, osteoblasts, osteogenesis, osteogenesis imperfecta, poly(A) tail, polyadenylation,
• MeSH
• buněčná diferenciace MeSH
• fyziologická kalcifikace genetika   MeSH
• kolagen typu I, řetězec alfa 1 genetika   metabolismus   MeSH
• kolagen typu I genetika   metabolismus   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši knockoutované MeSH
• myši MeSH
• neurotrofní faktory genetika   metabolismus   MeSH
• nukleotidyltransferasy genetika   metabolismus   MeSH
• oční proteiny genetika   metabolismus   MeSH
• osteoblasty metabolismus   patologie   MeSH
• osteogenesis imperfecta genetika   metabolismus   patologie   MeSH
• osteogeneze genetika   MeSH
• osteonektin genetika   metabolismus   MeSH
• poly(A)-polymerasa genetika   metabolismus   MeSH
• polyadenylace MeSH
• protein - isoformy nedostatek   genetika   MeSH
• sekvenční analýza RNA MeSH
• serpiny genetika   metabolismus   MeSH
• signální transdukce MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Col1a1 protein, mouse MeSH Prohlížeč
• Col1a2 protein, mouse MeSH Prohlížeč
• kolagen typu I, řetězec alfa 1 MeSH
• kolagen typu I MeSH
• messenger RNA MeSH
• neurotrofní faktory MeSH
• nukleotidyltransferasy MeSH
• oční proteiny MeSH
• osteonektin MeSH
• pigment epithelium-derived factor MeSH Prohlížeč
• poly(A)-polymerasa MeSH
• protein - isoformy MeSH
• serpiny MeSH
• SPARC protein, mouse MeSH Prohlížeč
• TENT5A protein, human MeSH Prohlížeč
• Tent5A protein, mouse MeSH Prohlížeč
• Tent5c protein, mouse MeSH Prohlížeč

Translocation of sterols between cellular membrane leaflets is of key importance in membrane organization, dynamics, and signaling. We present a novel translocation mechanism that differs in a unique manner from the established ones. The bobbing mechanism identified here is demonstrated for tail-oxidized sterols, but is expected to be viable for any molecule containing two polar centers at the opposite sides of the molecule. The mechanism renders translocation across a lipid membrane possible without a change in molecular orientation. For tail-oxidized sterols, the bobbing mechanism provides an exceptionally facile means to translocate these signaling molecules across membrane structures and may thus represent an important pathway in the course of their biological action.

• MeSH
• buněčná membrána chemie   metabolismus   MeSH
• cholesterol chemie   metabolismus   MeSH
• lipidové dvojvrstvy chemie   metabolismus   MeSH
• oxidace-redukce MeSH
• oxysteroly chemie   metabolismus   MeSH
• signální transdukce MeSH
• simulace molekulární dynamiky MeSH
• steroly chemie   metabolismus   MeSH
• termodynamika MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cholesterol MeSH
• lipidové dvojvrstvy MeSH
• oxysteroly MeSH
• steroly MeSH

An intact and functional sperm plasmalemma has a major role in sperm motility and fertilizing capacity. Several techniques have been developed to evaluate the integrity of the sperm plasma membrane, but there are still some inconsistencies concerning the methods that are more closely associated with sperm function. In this study, the aim was to: i) evaluate the integrity of the boar sperm plasmalemma during 72h of semen storage at 17°C using four techniques: eosin/nigrosin (E/N), propidium iodide/carboxyfluorescein diacetate (PI/CFDA), hypo-osmotic swelling test (HOST), and combined HOST with eosin staining (HOST/E), ii) assess the correlations and the limits of consistency among these techniques, iii) and estimate the relationships with the acrosomal status and sperm kinetics. Results indicate that the integrity of the sperm plasmalemma decreases during 72h of storage, although significant differences were found only using the HOST and HOST/E techniques. Moreover, use of E/N and PI/CFDA results in greater values relative to the undamaged sperm membrane than use of HOST and HOST/E at any incubation time. Overall, results using all techniques were consistent and correlate except for findings with PI/CFDA and HOST, which was slightly below 95%. Moreover, values using the techniques for the evaluation of the integrity of the sperm head and tail membranes are positively associated with the acrosomal status and different kinetic variables with the tail integrity being related to rapid linear trajectories and the head integrity to rapid curvilinear trajectories. The results of this study provide new insights into the relevance of evaluating the boar sperm plasmalemma in the routine spermiogram.

• Klíčová slova
• Pig, Plasma membrane, Semen storage, Sperm lifespan, Sperm velocity, Sperm viability,
• MeSH
• analýza spermatu veterinární   MeSH
• buněčná membrána MeSH
• časové faktory MeSH
• hlavička spermie MeSH
• motilita spermií * MeSH
• odběr biologického vzorku MeSH
• prasata MeSH
• spermie cytologie   fyziologie   MeSH
• teplota MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Escherichia coli phage SU10 belongs to the genus Kuravirus from the class Caudoviricetes of phages with short non-contractile tails. In contrast to other short-tailed phages, the tails of Kuraviruses elongate upon cell attachment. Here we show that the virion of SU10 has a prolate head, containing genome and ejection proteins, and a tail, which is formed of portal, adaptor, nozzle, and tail needle proteins and decorated with long and short fibers. The binding of the long tail fibers to the receptors in the outer bacterial membrane induces the straightening of nozzle proteins and rotation of short tail fibers. After the re-arrangement, the nozzle proteins and short tail fibers alternate to form a nozzle that extends the tail by 28 nm. Subsequently, the tail needle detaches from the nozzle proteins and five types of ejection proteins are released from the SU10 head. The nozzle with the putative extension formed by the ejection proteins enables the delivery of the SU10 genome into the bacterial cytoplasm. It is likely that this mechanism of genome delivery, involving the formation of the tail nozzle, is employed by all Kuraviruses.

• MeSH
• bakteriofágy * genetika   metabolismus   MeSH
• DNA virů genetika   MeSH
• fosmet * MeSH
• genom virový genetika   MeSH
• Podoviridae * genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA virů MeSH
• fosmet * MeSH

Cytoplasmic male sterility (CMS), encoded by the interacting mitochondrial and nuclear genes, causes pollen abortion or non-viability. CMS is widely used in agriculture and extensively studied in crops. Much less is known about CMS in wild species. We performed a comparative transcriptomic analysis of male sterile and fertile individuals of Silene vulgaris, a model plant for the study of gynodioecy, to reveal the genes responsible for pollen abortion in this species. We used RNA-seq datasets previously employed for the analysis of mitochondrial and plastid transcriptomes of female and hermaphrodite flower buds, making it possible to compare the transcriptomes derived from three genomes in the same RNA specimen. We assembled de novo transcriptomes for two haplotypes of S. vulgaris and identified differentially expressed genes between the females and hermaphrodites, associated with stress response or pollen development. The gene for alternative oxidase was downregulated in females. The genetic pathways controlling CMS in S. vulgaris are similar to those in crops. The high number of the differentially expressed nuclear genes contrasts with the uniformity of organellar transcriptomes across genders, which suggests these pathways are evolutionarily conserved and that selective mechanisms may shield organellar transcription against changes in the cytoplasmic transcriptome.

• Klíčová slova
• Silene vulgaris, cytoplasmic male sterility, differential gene expression,
• MeSH
• anotace sekvence MeSH
• buněčné jádro genetika   MeSH
• down regulace genetika   MeSH
• genová ontologie MeSH
• haplotypy genetika   MeSH
• messenger RNA genetika   metabolismus   MeSH
• mitochondriální proteiny genetika   metabolismus   MeSH
• neplodnost rostlin genetika   MeSH
• oxidační stres genetika   MeSH
• oxidoreduktasy genetika   metabolismus   MeSH
• pyl genetika   MeSH
• regulace genové exprese u rostlin * MeSH
• rostlinné proteiny genetika   metabolismus   MeSH
• Silene genetika   fyziologie   MeSH
• stanovení celkové genové exprese * MeSH
• transkriptom genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alternative oxidase MeSH Prohlížeč
• messenger RNA MeSH
• mitochondriální proteiny MeSH
• oxidoreduktasy MeSH
• rostlinné proteiny MeSH

Retroviral envelope glycoprotein (Env) is essential for the specific recognition of the host cell and the initial phase of infection. As reported for human immunodeficiency virus (HIV), the recruitment of Env into a retroviral membrane envelope is mediated through its interaction with a Gag polyprotein precursor of structural proteins. This interaction, occurring between the matrix domain (MA) of Gag and the cytoplasmic tail (CT) of the transmembrane domain of Env, takes place at the host cell plasma membrane. To determine whether the MA of Mason-Pfizer monkey virus (M-PMV) also interacts directly with the CT of Env, we mimicked the in vivo conditions in an in vitro experiment by using a CT in its physiological trimeric conformation mediated by the trimerization motif of the GCN4 yeast transcription factor. The MA protein was used at the concentration shifting the equilibrium to its trimeric form. The direct interaction between MA and CT was confirmed by a pulldown assay. Through the combination of nuclear magnetic resonance (NMR) spectroscopy and protein cross-linking followed by mass spectrometry analysis, the residues involved in mutual interactions were determined. NMR has shown that the C terminus of the CT is bound to the C-terminal part of MA. In addition, protein cross-linking confirmed the close proximity of the N-terminal part of CT and the N terminus of MA, which is enabled in vivo by their location at the membrane. These results are in agreement with the previously determined orientation of MA on the membrane and support the already observed mechanisms of M-PMV virus-like particle transport and budding.IMPORTANCE By a combination of nuclear magnetic resonance (NMR) and mass spectroscopy of cross-linked peptides, we show that in contrast to human immunodeficiency virus type 1 (HIV-1), the C-terminal residues of the unstructured cytoplasmic tail of Mason-Pfizer monkey virus (M-PMV) Env interact with the matrix domain (MA). Based on biochemical data and molecular modeling, we propose that individual cytoplasmic tail (CT) monomers of a trimeric complex bind MA molecules belonging to different neighboring trimers, which may stabilize the MA orientation at the membrane by the formation of a membrane-bound net of interlinked Gag and CT trimers. This also corresponds with the concept that the membrane-bound MA of Gag recruits Env through interaction with the full-length CT, while CT truncation during maturation attenuates the interaction to facilitate uncoating. We propose a model suggesting different arrangements of MA-CT complexes between a D-type and C-type retroviruses with short and long CTs, respectively.

• Klíčová slova
• Env, M-PMV, Retrovirus, cytoplasmic tail, protein interactions,
• MeSH
• genové produkty env chemie   genetika   MeSH
• genové produkty gag chemie   genetika   MeSH
• Masonův-Pfizerův opičí virus chemie   genetika   MeSH
• proteinové domény MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• genové produkty env MeSH
• genové produkty gag MeSH

Fonticins are phage tail-like bacteriocins produced by the Gram-negative bacterium Pragia fontium from the family Budviciaceae. This bacterium produces contractile-type particles that adsorb on the surface of sensitive bacteria and penetrate the cell wall, probably during contraction, in a way similar to the type VI secretion system. We characterized the pore-forming activity of fonticins using both living cells and in vitro model membranes. Using a potassium leakage assay, we show that fonticins are able to permeabilize sensitive cells. On black lipid membranes, single-pore conductance is about 0.78 nS in 1 M NaCl and appears to be linearly dependent on the increasing molar strength of NaCl solution, which is a property of considerably large pores. In agreement with these findings, fonticins are not ion selective for Na+, K+, and Cl-. Polyethylene glycol 3350 (PEG 3350) molecules of about 3.5 nm in diameter can enter the fonticin pore lumen, whereas the larger molecules cannot pass the pore. The size of fonticin pores was confirmed by transmission electron microscopy. The terminal membrane-piercing complex of the fonticin tube probably creates a selective barrier restricting passage of macromolecules. IMPORTANCE Phage tail-like bacteriocins are now the subject of research as potent antibacterial agents due to their narrow host specificity and single-hit mode of action. In this work, we focused on the structure and mode of action of fonticins. According to some theories, related particles were initially adapted for passage of double-stranded DNA (dsDNA) molecules, but fonticins changed their function during the evolution; they are able to form large pores through the bacterial envelope of Gram-negative bacteria. As various pore-forming proteins are extensively used for nanopore sequencing and stochastic sensing, we decided to investigate the pore-forming properties of fonticin protein complexes on artificial lipid membranes. Our research revealed remarkable structural properties of these particles that may have a potential application as a nanodevice.

• Klíčová slova
• black lipid membranes, conductance, electric current, electron microscopy, fonticin, membrane pore formation, phage tail-like bacteriocins, single-pore conductance,
• MeSH
• bakteriociny * metabolismus   MeSH
• buněčná membrána metabolismus   MeSH
• chlorid sodný metabolismus   MeSH
• Enterobacteriaceae MeSH
• Gammaproteobacteria MeSH
• lipidové dvojvrstvy * metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriociny * MeSH
• chlorid sodný MeSH
• lipidové dvojvrstvy * MeSH