POSITION OF MICRO AND SMALL ENTERPRISES LIMITED LIABILITY COMPANY REVIEWED FROM LAW NUMBER 11 OF 2020 CONCERNING WORK CREATION

This study aims to determine the position of Micro and Small Enterprises Limited Liability Company as regulated in Law Number 11 of 2020 concerning of Job Creation, as well as the way of its establishment and the form of responsibility for the founders of Micro and Small Enterprises Limited Liability Company. This research is a type of normative legal research using a statutory approach method and a conceptual approach. From the results of the study it is concluded that first, the position of Micro and Small Enterprises Limited Liability Company is as an individual legal entity where this Limited Liability Company must reach the criteria for micro and small businesses as stipulated in Government Regulation Number 7 of 2021. Second, the method of establishing Micro and Small Enterprises Limited Liability Company, to Private Company was established based on the provisions of the Job Creation Law and Government Regulation Number 8 of 2021, while for the Company it was established based on the provisions of Law Number 40 of 2008 concerning Limited Liability Companies. Third, the form of responsibility of the founder of Micro and Small Enterprises Limited Liability Company, both before and after being legalized as a legal entity, is the same as the form of responsibility of the Limited Liability Company in general. Keywords: Position, Micro and Small Enterprises Limited Liability Company DOI: 10.7176/JLPG/108-10 Publication date: April 30th 202

Geosmithia argillacea: an emerging agent of airway colonization in patients with cystic fibrosis?

Date du colloque&nbsp;: 06/2009</p

Wnt signaling is boosted during intestinal regeneration by a CD44-positive feedback loop

Enhancement of Wnt signaling is fundamental for stem cell function during intestinal regeneration. Molecular modules control Wnt activity by regulating signal transduction. CD44 is such a positive regulator and a Wnt target gene. While highly expressed in intestinal crypts and used as a stem cell marker, its role during intestinal homeostasis and regeneration remains unknown. Here we propose a CD44 positive-feedback loop that boosts Wnt signal transduction, thus impacting intestinal regeneration. Excision of Cd44 in Cd44$^{fl/fl}$;VillinCreER$^{T2}$ mice reduced Wnt target gene expression in intestinal crypts and affected stem cell functionality in organoids. Although the integrity of the intestinal epithelium was conserved in mice lacking CD44, they were hypersensitive to dextran sulfate sodium, and showed more severe inflammation and delayed regeneration. We localized the molecular function of CD44 at the Wnt signalosome, and identified novel DVL/CD44 and AXIN/CD44 complexes. CD44 thus promotes optimal Wnt signaling during intestinal regeneration

Nucleic Acids Res

The HIV-1 nucleocapsid protein (NCp7) is a nucleic acid chaperone required during reverse transcription. During the first strand transfer, NCp7 is thought to destabilize cTAR, the (-)DNA copy of the TAR RNA hairpin, and subsequently direct the TAR/cTAR annealing through the zipping of their destabilized stem ends. To further characterize the destabilizing activity of NCp7, we locally probe the structure and dynamics of cTAR by steady-state and time resolved fluorescence spectroscopy. NC(11-55), a truncated NCp7 version corresponding to its zinc-finger domain, was found to bind all over the sequence and to preferentially destabilize the penultimate double-stranded segment in the lower part of the cTAR stem. This destabilization is achieved through zinc-finger-dependent binding of NC to the G(10) and G(50) residues. Sequence comparison further revealed that C*A mismatches close to the two G residues were critical for fine tuning the stability of the lower part of the cTAR stem and conferring to G(10) and G(50) the appropriate mobility and accessibility for specific recognition by NC. Our data also highlight the necessary plasticity of NCp7 to adapt to the sequence and structure variability of cTAR to chaperone its annealing with TAR through a specific pathway

Intrinsic nucleic acid dynamics modulates HIV-1 nucleocapsid protein binding to its targets

HIV-1 nucleocapsid protein (NC) is involved in the rearrangement of nucleic acids occurring in key steps of reverse transcription. The protein, through its two zinc fingers, interacts preferentially with unpaired guanines in single-stranded sequences. In mini-cTAR stem-loop, which corresponds to the top half of the cDNA copy of the transactivation response element of the HIV-1 genome, NC was found to exhibit a clear preference for the TGG sequence at the bottom of mini-cTAR stem. To further understand how this site was selected among several potential binding sites containing unpaired guanines, we probed the intrinsic dynamics of mini-cTAR using (13)C relaxation measurements. Results of spin relaxation time measurements have been analyzed using the model-free formalism and completed by dispersion relaxation measurements. Our data indicate that the preferentially recognized guanine in the lower part of the stem is exempt of conformational exchange and highly mobile. In contrast, the unrecognized unpaired guanines of mini-cTAR are involved in conformational exchange, probably related to transient base-pairs. These findings support the notion that NC preferentially recognizes unpaired guanines exhibiting a high degree of mobility. The ability of NC to discriminate between close sequences through their dynamic properties contributes to understanding how NC recognizes specific sites within the HIV genome

PloS one

The HIV-1 nucleocapsid protein (NC) is a small basic protein containing two zinc fingers (ZF) separated by a short linker. It is involved in several steps of the replication cycle and acts as a nucleic acid chaperone protein in facilitating nucleic acid strand transfers occurring during reverse transcription. Recent analysis of three-dimensional structures of NC-nucleic acids complexes established a new property: the unpaired guanines targeted by NC are more often inserted in the C-terminal zinc finger (ZF2) than in the N-terminal zinc finger (ZF1). Although previous NMR dynamic studies were performed with NC, the dynamic behavior of the linker residues connecting the two ZF domains remains unclear. This prompted us to investigate the dynamic behavior of the linker residues. Here, we collected 15N NMR relaxation data and used for the first time data at several fields to probe the protein dynamics. The analysis at two fields allows us to detect a slow motion occurring between the two domains around a hinge located in the linker at the G35 position. However, the amplitude of motion appears limited in our conditions. In addition, we showed that the neighboring linker residues R29, A30, P31, R32, K33 displayed restricted motion and numerous contacts with residues of ZF1. Our results are fully consistent with a model in which the ZF1-linker contacts prevent the ZF1 domain to interact with unpaired guanines, whereas the ZF2 domain is more accessible and competent to interact with unpaired guanines. In contrast, ZF1 with its large hydrophobic plateau is able to destabilize the double-stranded regions adjacent to the guanines bound by ZF2. The linker residues and the internal dynamics of NC regulate therefore the different functions of the two zinc fingers that are required for an optimal chaperone activity

Rsc Adv

A Forster resonance energy transfer (FRET) system of semiconductor quantum dots and porphyrins represents a new promising photosensitizing tool for the photodynamic therapy of cancer. In this work, we demonstrate the ability of a non-covalent complex formed between commercial lipid-coated CdSe/ ZnS quantum dots (QD) bearing different terminal groups (carboxyl, amine or non-functionalized) and a second-generation photosensitizer, chlorin e(6) (Ce-6) to enter living HeLa cells with maintained integrity and perform FRET from two-photon excited QD to bound Ce-6 molecules. Spectroscopic changes, the highly efficient FRET, observed upon Ce-6 binding to QD, and remarkable stability of the QD-Ce-6 complex in different media suggest that Ce-6 penetrates inside the lipid coating close to the inorganic core of QD. Two-photon fluorescence lifetime imaging microscopy (FLIM) on living HeLa cells revealed that QD-Ce-6 complexes localize within the plasma membrane and intracellular compartments and preserve high FRET efficiency (similar to 50%). The latter was confirmed by recovery of QD emission lifetime after photobleaching of Ce-6. The intracellular distribution pattern and FRET efficiency of QD-Ce-6 complexes did not depend on the charge of QD terminal groups. Given the non-covalent nature of the complex, its exceptional stability in cellulo can be explained by a combination of hydrophobic interactions and coordination of carboxyl groups of Ce6 with the ZnS shell of QD. These findings suggest a simple route to the preparation of QD-photosensitizer complexes featuring efficient FRET and high stability in cellulo without using time-consuming conjugation protocols