scholarly journals Uncovering the distinct properties of a bacterial type I-E CRISPR activation system

2021 ◽  
Author(s):  
Maria Claudia Villegas Kcam ◽  
Annette J. Tsong ◽  
James Chappell
Keyword(s):  
Type I ◽  
Type Ii ◽  
Activation Patterns ◽  
Activation Domains ◽  
Control Gene Expression ◽  
Activation System ◽  
Alternative Type ◽  
Regulatory Properties ◽  
Bacterial Type ◽  
Crispr Activation

ABSTRACTSynthetic gene regulators based upon CRISPR-Cas systems offer highly programmable technologies to control gene expression in bacteria. Bacterial CRISPR activators (CRISPRa) have been developed that use engineered type II CRISPR-dCas9 to localize transcription activation domains near promoter elements to activate transcription. However, several reports have demonstrated distance-dependent requirements and periodical activation patterns that overall limit the flexibility of these systems. Here, we demonstrate the potential of using an alternative type I-E CRISPR-Cas system to create a CRISPRa with distinct and expanded regulatory properties. We create the first bacterial CRISPRa system based upon a type I-E CRISPR-Cas, and demonstrate differences in the activation range of this system compared to type II CRISPRa systems. Furthermore, we characterize the distance-dependent activation patterns of type I-E CRISPRa to reveal a distinct and more frequent periodicity of activation.

scholarly journals Electromyographic activation patterns during swallowing in older adults

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jin Young Ko ◽  
Hayoung Kim ◽  
Joonyoung Jang ◽  
Jun Chang Lee ◽  
Ju Seok Ryu
Keyword(s):  
Older Adults ◽  
Viscous Fluid ◽  
Muscle Activation ◽  
Fatty Infiltration ◽  
Liquid Volume ◽  
Type I ◽  
Type Ii ◽  
Activation Patterns ◽  
Muscle Activation Patterns ◽  
Age Related

AbstractAge-related weakness due to atrophy and fatty infiltration in oropharyngeal muscles may be related to dysphagia in older adults. However, little is known about changes in the oropharyngeal muscle activation pattern in older adults. This was a prospective and experimental study. Forty healthy participants (20 older [> 60 years] and 20 young [< 60 years] adults) were enrolled. Six channel surface electrodes were placed over the bilateral suprahyoid (SH), bilateral retrohyoid (RH), thyrohyoid (TH), and sternothyroid (StH) muscles. Electromyography signals were then recorded twice for each patient during swallowing of 2 cc of water, 5 cc of water, and 5 cc of a highly viscous fluid. Latency, duration, and peak amplitude were measured. The activation patterns were the same, in the order of SH, TH, and StH, in both groups. The muscle activation patterns were classified as type I and II; the type I pattern was characterized by a monophasic shape, and the type II comprised a pre-reflex phase and a main phase. The oropharyngeal muscles and SH muscles were found to develop a pre-reflex phase specifically with increasing volume and viscosity of the swallowed fluid. Type I showed a different response to the highly viscous fluid in the older group compared to that in the younger group. However, type II showed concordant changes in the groups. Therefore, healthy older people were found to compensate for swallowing with a pre-reflex phase of muscle activation in response to increased liquid volume and viscosity, to adjust for age-related muscle weakness.

scholarly journals Genetic basis of the formation and identity of type I and type II neurons in Drosophila embryos

Development ◽  
1997 ◽  
Vol 124 (14) ◽  
pp. 2819-2828 ◽  
Author(s):  
M. Vervoort ◽  
D.J. Merritt ◽  
A. Ghysen ◽  
C. Dambly-Chaudiere
Keyword(s):  
Cell Communication ◽  
Sense Organ ◽  
Type I ◽  
Type Ii ◽  
Neuronal Fate ◽  
Late Step ◽  
Alternative Type ◽  
The Difference ◽  
Dependent Lineages ◽  
Multidendritic Neurons

The embryonic peripheral nervous system of Drosophila contains two main types of sensory neurons: type I neurons, which innervate external sense organs and chordotonal organs, and type II multidendritic neurons. Here, we analyse the origin of the difference between type I and type II in the case of the neurons that depend on the proneural genes of the achaete-scute complex (ASC). We show that, in Notch- embryos, the type I neurons are missing while type II neurons are produced in excess, indicating that the type I/type II choice relies on Notch-mediated cell communication. In contrast, both type I and type II neurons are absent in numb- embryos and after ubiquitous expression of tramtrack, indicating that the activity of numb and the absence of tramtrack are required to produce both external sense organ and multidendritic neural fates. The analysis of string- embryos reveals that when the precursors are unable to divide they differentiate mostly into type II neurons, indicating that the type II is the default neuronal fate. We also report a new mutant phenotype where the ASC-dependent neurons are converted into type II neurons, providing evidence for the existence of one or more genes required for maintaining the alternative (type I) fate. Our results suggest that the same mechanism of type I/type II specification may operate at a late step of the ASC-dependent lineages, when multidendritic neurons arise as siblings of the external sense organ neurons and, at an early step, when other multidendritic neurons precursors arise as siblings of external sense organ precursors.

scholarly journals Ungeremine effectively targets mammalian as well as bacterial type I and type II topoisomerases

2011 ◽  
Vol 21 (23) ◽  
pp. 7041-7044 ◽  
Author(s):  
Laura Casu ◽  
Filippo Cottiglia ◽  
Marco Leonti ◽  
Alessandro De Logu ◽  
Emanuela Agus ◽  
...  
Keyword(s):  
Type I ◽  
Type Ii ◽  
Bacterial Type

scholarly journals Pyocin S2 (Sa) Kills Pseudomonas aeruginosa Strains via the FpvA Type I Ferripyoverdine Receptor

2007 ◽  
Vol 189 (21) ◽  
pp. 7663-7668 ◽  
Author(s):  
Sarah Denayer ◽  
Sandra Matthijs ◽  
Pierre Cornelis
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Type I ◽  
Type Ii ◽  
Specific Receptor ◽  
Immunity Protein ◽  
Immunity Gene ◽  
Gene Comparison ◽  
Signal Type ◽  
Alternative Type ◽  
Resistant Strains

ABSTRACT Soluble (S-type) pyocins are Pseudomonas aeruginosa bacteriocins that kill nonimmune P. aeruginosa strains via a specific receptor. The genes coding for pyocin Sa (consisting of a killing protein and an immunity protein) were cloned and expressed in Escherichia coli. Sequence analysis revealed that Sa is identical to pyocin S2. Seventy-nine strains of P. aeruginosa were tested for their sensitivity to pyocins S1, S2, and S3, and their ferripyoverdine receptors were typed by multiplex PCR. No strain was found to be sensitive to both S2 and S3, suggesting that the receptors for these two pyocins cannot coexist in one strain. As expected, all S3-sensitive strains had the type II ferripyoverdine receptor fpvA gene, confirming our previous reports. S1 killed strains irrespective of the type of ferripyoverdine receptor they produced. All S2-sensitive strains had the type I fpvA gene, and the inactivation of type I fpvA in an S2-sensitive strain conferred resistance to the S2 pyocin. Accordingly, complementation with type I fpvA in trans restored sensitivity to S2. Some S2-resistant type I fpvA-positive strains were detected, the majority (all but five) of which had the S1-S2 immunity gene. Comparison of type I fpvA sequences from immunity gene-negative S2-sensitive and S2-resistant strains revealed only a valine-to-isoleucine substitution at position 46 of type I FpvA. However, both type I fpvA genes conferred the capacity for type I pyoverdine utilization and sensitivity to S2. When these two type I fpvA genes were introduced into strain 7NSK2 carrying mutations in type II fpvA (encoding the type II pyoverdine receptor) and fpvB (encoding the alternative type I receptor), growth in the presence of type I pyoverdine was observed and the strain became sensitive to S2. We also found that type I pyoverdine could signal type II pyoverdine production via the type I FpvA receptor in 7NSK2.

Chromosomal bacterial type II toxin–antitoxin systems

2012 ◽  
Vol 58 (5) ◽  
pp. 553-562 ◽  
Author(s):  
Mohammad Adnan Syed ◽  
Céline M. Lévesque
Keyword(s):  
De Novo ◽  
Physiological Role ◽  
Type I ◽  
Type Ii ◽  
Translation Process ◽  
Bacterial Physiology ◽  
The Subject ◽  
Almost All ◽  
Bacterial Type

Most prokaryotic chromosomes contain a number of toxin–antitoxin (TA) modules consisting of a pair of genes that encode 2 components, a stable toxin and its cognate labile antitoxin. TA systems are also known as addiction modules, since the cells become “addicted” to the short-lived antitoxin product (the unstable antitoxin is degraded faster than the more stable toxin) because its de novo synthesis is essential for their survival. While toxins are always proteins, antitoxins are either RNAs (type I, type III) or proteins (type II). Type II TA systems are widely distributed throughout the chromosomes of almost all free-living bacteria and archaea. The vast majority of type II toxins are mRNA-specific endonucleases arresting cell growth through the mechanism of RNA cleavage, thus preventing the translation process. The physiological role of chromosomal type II TA systems still remains the subject of debate. This review describes the currently known type II toxins and their characteristics. The different hypotheses that have been proposed to explain their role in bacterial physiology are also discussed.

scholarly journals Overlapping, Additive and Counterregulatory Effects of Type II and I Interferons on Myeloid Dendritic Cell Functions

2011 ◽  
Vol 11 ◽  
pp. 2071-2090 ◽  
Author(s):  
Loredana Frasca ◽  
Roberto Lande
Keyword(s):  
Recent Literature ◽  
Type I ◽  
Type Ii ◽  
Vaccine Adjuvants ◽  
Therapeutic Approaches ◽  
Innate And Adaptive Immunity ◽  
Therapeutic Implications ◽  
Cell Functions ◽  
Type I Ifns ◽  
Regulatory Properties

Dendritic cells (DCs) are central player in immunity by bridging the innate and adaptive arms of the immune system (IS). Interferons (IFNs) are one of the most important factors that regulate both innate and adaptive immunity too. Thus, the understanding of how type II and I IFNs modulate the immune-regulatory properties of DCs is a central issue in immunology. In this paper, we will address this point in the light of the most recent literature, also highlighting the controversial data reported in the field. According to the wide literature available, type II as well as type I IFNs appear, at the same time, to collaborate, to induce additive effects or overlapping functions, as well as to counterregulate each one's effects on DC biology and, in general, the immune response. The knowledge of these effects has important therapeutic implications in the treatment of infectious/autoimmune diseases and cancer and indicates strategies for using IFNs as vaccine adjuvants and in DC-based immune therapeutic approaches.

scholarly journals Comparative interactomics analysis reveals potential regulators of α6β4 distribution in keratinocytes

Biology Open ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. bio054155
Author(s):  
Lisa te Molder ◽  
Liesbeth Hoekman ◽  
Maaike Kreft ◽  
Onno Bleijerveld ◽  
Arnoud Sonnenberg
Keyword(s):  
Close Association ◽  
Cortical Microtubule ◽  
Focal Adhesions ◽  
Type I ◽  
Type Ii ◽  
Essential Components ◽  
Integrin Α6β4 ◽  
Alternative Type ◽  
Labeling Approach ◽  
Integrin Α3β1

ABSTRACTThe integrin α6β4 and cytoskeletal adaptor plectin are essential components of type I and type II hemidesmosomes (HDs). We recently identified an alternative type II HD adhesion complex that also contains CD151 and the integrin α3β1. Here, we have taken a BioID proximity labeling approach to define the proximity protein environment for α6β4 in keratinocytes. We identified 37 proteins that interacted with both α6 and β4, while 20 and 78 proteins specifically interacted with the α6 and β4 subunits, respectively. Many of the proximity interactors of α6β4 are components of focal adhesions (FAs) and the cortical microtubule stabilizing complex (CMSC). Though the close association of CMSCs with α6β4 in HDs was confirmed by immunofluorescence analysis, CMSCs have no role in the assembly of HDs. Analysis of the β4 interactome in the presence or absence of CD151 revealed that they are strikingly similar; only 11 different interactors were identified. One of these was the integrin α3β1, which interacted with α6β4 more strongly in the presence of CD151 than in its absence. These findings indicate that CD151 does not significantly contribute to the interactome of α6β4, but suggest a role of CD151 in linking α3β1 and α6β4 together in tetraspanin adhesion structures.

scholarly journals EPR Spectroscopy of MolB2C2-A Reveals Mechanism of Transport for a Bacterial Type II Molybdate Importer

2013 ◽  
Vol 288 (29) ◽  
pp. 21228-21235 ◽  
Author(s):  
Austin J. Rice ◽  
Frances J. D. Alvarez ◽  
Kathryn M. Schultz ◽  
Candice S. Klug ◽  
Amy L. Davidson ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Abc Transporters ◽  
Epr Spectroscopy ◽  
Vitamin B ◽  
Type I ◽  
Type Ii ◽  
Bacterial Type ◽  
Uptake Of Nutrients ◽  
Structure And Activity ◽  
Alternating Access

In bacteria, ATP-binding cassette (ABC) transporters are vital for the uptake of nutrients and cofactors. Based on differences in structure and activity, ABC importers are divided into two types. Type I transporters have been well studied and employ a tightly regulated alternating access mechanism. Less is known about Type II importers, but much of what we do know has been observed in studies of the vitamin B12 importer BtuC2D2. MolB2C2 (formally known as HI1470/71) is also a Type II importer, but its substrate, molybdate, is ∼10-fold smaller than vitamin B12. To understand mechanistic differences among Type II importers, we focused our studies on MolBC, for which alternative conformations may be required to transport its relatively small substrate. To investigate the mechanism of MolBC, we employed disulfide cross-linking and EPR spectroscopy. From these studies, we found that nucleotide binding is coupled to a conformational shift at the periplasmic gate. Unlike the larger conformational changes in BtuCD-F, this shift in MolBC-A is akin to unlocking a swinging door: allowing just enough space for molybdate to slip into the cell. The lower cytoplasmic gate, identified in BtuCD-F as “gate I,” remains open throughout the MolBC-A mechanism, and cytoplasmic gate II closes in the presence of nucleotide. Combining our results, we propose a peristaltic mechanism for MolBC-A, which gives new insight in the transport of small substrates by a Type II importer.

Heat-Etching with a Bullivant Type II Simple Freeze-Cleave Device

1970 ◽  
Vol 28 ◽  
pp. 106-107 ◽  
Author(s):  
Ronald S. Weinstein ◽  
N. Scott McNutt
Keyword(s):  
New Zealand ◽  
General Hospital ◽  
Massachusetts General Hospital ◽  
Type I ◽  
Type Ii ◽  
Collaborative Effort ◽  
Temperature And Pressure ◽  
The University

The Type I simple cold block device was described by Bullivant and Ames in 1966 and represented the product of the first successful effort to simplify the equipment required to do sophisticated freeze-cleave techniques. Bullivant, Weinstein and Someda described the Type II device which is a modification of the Type I device and was developed as a collaborative effort at the Massachusetts General Hospital and the University of Auckland, New Zealand. The modifications reduced specimen contamination and provided controlled specimen warming for heat-etching of fracture faces. We have now tested the Mass. General Hospital version of the Type II device (called the “Type II-MGH device”) on a wide variety of biological specimens and have established temperature and pressure curves for routine heat-etching with the device.

Labelling of non-A, non-B hepatitis infected chimpanzee hepatocytes with nuclease-gold conjugates

1984 ◽  
Vol 42 ◽  
pp. 250-251
Author(s):  
G. D. Gagne ◽  
M. F. Miller ◽  
D. A. Peterson
Keyword(s):  
Endoplasmic Reticulum ◽  
Experimental Infection ◽  
Uranyl Acetate ◽  
Type I ◽  
Cellular Injury ◽  
Type Ii ◽  
Tubular Structures ◽  
Type Iii ◽  
Type Iv ◽  
Infected Chimpanzee

Experimental infection of chimpanzees with non-A, non-B hepatitis (NANB) or with delta agent hepatitis results in the appearance of characteristic cytoplasmic alterations in the hepatocytes. These alterations include spongelike inclusions (Type I), attached convoluted membranes (Type II), tubular structures (Type III), and microtubular aggregates (Type IV) (Fig. 1). Type I, II and III structures are, by association, believed to be derived from endoplasmic reticulum and may be morphogenetically related. Type IV structures are generally observed free in the cytoplasm but sometimes in the vicinity of type III structures. It is not known whether these structures are somehow involved in the replication and/or assembly of the putative NANB virus or whether they are simply nonspecific responses to cellular injury. When treated with uranyl acetate, type I, II and III structures stain intensely as if they might contain nucleic acids. If these structures do correspond to intermediates in the replication of a virus, one might expect them to contain DNA or RNA and the present study was undertaken to explore this possibility.

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