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SecA facilitates bacterial protein translocation by its association with presecretory or membrane proteins and the Sec EG translocon channel. Once assembled, SecA ATPase undergoes cycles of membrane insertion and retraction at SecYEG that drive protein translocation in a stepwise fashion. SecA exists in equilibrium between a monomer and dimer, and association with its translocation ligands shifts this equilibrium dramatically. Here, we examined the proposal that protein translocation can occur by means of a SecA monomer. We produced a mutant SecA protein lacking residues 2-11, which was found to exist mostly as a monomer, and it was unable to complement a conditional-lethal secA mutant was inactive for in vitro protein translocation, and was poorly active for translocation ATPase activity. Furthermore, we developed a technique termed membrane trapping, where wild-type Seth subunits became trapped within the membrane by overproduction of membrane-stuck mutant SecA proteins, and, in one case, a membrane- associated Seth heterodimer was demonstrated. Finally, we ex- amined both endogenous and reconstituted membrane-bound SecA and found a significant level of Seth dimer in both cases, as assessed by chemical crosslinking. Collectively, our results strongly suggest that membrane-bound SecA dimer is critical for the protein translocation cycle, although these results cannot exclude participation of Seth monomer at some stage in the translocation process. Our findings have important implications regarding SecA motor function and translocon assembly and activation. [ABSTRACT FROM AUTHOR]

Seca is an ATPase and motor protein that drives protein translocation across the bacterial plasma membrane. In Escherichia coli Seca levels are regulated by the secretion needs of the cell utilizing secM, which encodes a secreted protein. Previous studies demonstrated that this regulation requires a translational pause within secM, whose duration regulates the accessibility of the seca Shine-Dalgarno sequence on secM seca mRNA. Here we provide evidence that translocon "pulling" of nascent SecM is what regulates the duration of the secM translational pause, and thus seca expression levels, thereby providing direct support for this model. [ABSTRACT FROM AUTHOR]

Studies conformational changes induced by the lipid and signal peptide within the C-domain of Escherichia coli SecA protein using fluorescence spectroscopy. Translocation of preprotein segments to the trans side of the membrane; Role of integral membrane accessory proteins in polypeptide chain translocation; Conformational changes of proteins during the protein folding process.

The bacterial Sec-dependent system is the major protein-biogenic pathway for protein secretion across the cytoplasmic membrane or insertion of integral membrane proteins into the phospholipid bilayer. The mechanism of SecA-driven protein transport across the SecYEG channel complex has remained controversial with conflicting claims from biochemical and structural studies regarding the depth and extent of SecA insertion into SecYEG during ongoing protein transport. Here we utilized site-specific in vivo photo-crosslinking to thoroughly map SecY regions that are in contact with SecA during its insertion cycle. An arabinose-inducible, rapidly folding OmpA-GFP chimera was utilized to jam the SecYEG channels with an arrested substrate protein to "freeze" them in their SecA-inserted state. Examination of 117 sites distributed throughout SecY indicated that SecA not only interacts extensively with the cytosolic regions of SecY as shown previously, but it also interacts with most of the transmembrane helices and periplasmic regions of SecY, with a clustering of interaction sights around the lateral gate and pore ring regions. Our observations support previous reports of SecA membrane insertion during in vitro protein transport as well as those documenting the membrane penetration properties of this protein. They suggest that one or more SecA regions transiently integrate into the heart of the SecY channel complex to span the membrane to promote the protein transport cycle. These findings indicate that high-resolution structural information about the membrane-inserted state of SecA is still lacking and will be critical for elucidating the bacterial protein transport mechanism. [ABSTRACT FROM AUTHOR]

SecYEG protein of bacteria or Sec61α β γ of eukaryotes is a universally conserved heterotrimeric protein channel complex that accommodates the partitioning of membrane proteins into the lipid bilayer as well as the secretion of proteins to the trans side of the plasma or endoplasmic reticular membrane, respectively. SecYEG function is facilitated by cytosolic partners, mainly a nascent chain-ribosome complex or the SecA ATPase motor protein. Extensive efforts utilizing both biochemical and biophysical approaches have been made to determine whether SecYEG functions as a monomer or a dimer, but such approaches have often generated conflicting results. Here we have employed site-specific in vivo photo-cross-linking or cysteine cross-linking, along with co-immunoprecipitation or SecA footprinting techniques to readdress this issue. Our findings show that the SecY dimer to monomer ratio is relatively constant regardless of whether translocons are actively engaged with protein substrate or not. Under the former conditions the SecY dimer can be captured associated with a translocon-jammed substrate, indicative of SecY dimer function. Furthermore, SecA ATPase can be cross-linked to two copies of SecY when the complex contains a translocation intermediate. Collectively, our results suggest that SecYEG dimers are functional units of the translocon. [ABSTRACT FROM AUTHOR]

The SecAΔ nanomotor promotes protein translocation in eubacteria by binding both protein cargo and the protein-conducting channel and by undergoing ATP-driven conformation cycles that drive this process. There are conflicting reports about whether SecA functions as a monomer or dimer during this dynamic process. Here we reexamined the roles of the amino and carboxyl termini of SecAΔ in promoting its dimerization and functional state by examining three secA mutants and the corresponding proteins: SecAΔ8 lacking residues 2 to 8, SecAΔΔ11 lacking residues 2 to 11, and SecAΔ11/N95 lacking both residues 2 to 11 and the carboxyl-terminal 70 residues. We demonstrated that whether SecAΔ11 or SecAΔ11/N95 was functional for promoting cell growth depended solely on the vivo level of the protein, which appeared to govern residual dimerization. All three SecAΔ mutant proteins were defective for promoting cell growth unless they were highly overproduced. Cell fractionation revealed that SecAΔ11 and SecAΔ11/N95 were proficient in membrane association, although the formation of integral membrane SecAΔ was reduced. The presence of a modestly higher level of SecAΔ11/N95 in the membrane and the ability of this protein to form dimers, as detected by chemical cross-linking, were consistent with the higher level of SecAΔ expression and better growth of the SecAΔ11/N95 mutant than of the SecAΔ11 mutant. Biochemical studies showed that SecAΔ11 and SecAΔ11/N95 had identical dimerization defects, while SecAΔ8 was intermediate between these proteins and wild-type SecAΔ in terms of dimer formation. Furthermore, both SecAΔ11 and SecAΔ11/N95 were equally defective in translocation ATPase specific activity. Our studies showed that the nonessential carboxyl-terminal 70 residues of SecA play no role in its dimerization, while increasing the truncation of the amino-terminal region of SecA from 8 to 11 residues results in increased defects in SecA dimerization and poor in vivo function unless the protein is highly overexpressed. They also clarified a number of conflicting previous reports and support the essential nature of the SecA dimer. [ABSTRACT FROM AUTHOR]

The SecA adenosine triphosphatase (ATPase) mediates extrusion of the amino termini of secreted proteins from the eubacterial cytosol based on cycles of reversible binding to the SecYEG translocon. We have determined the crystal structure of SecA with and without magnesium—adenosine diphosphate bound to the high-affinity ATPase site at 3.0 and 2.7 angstrom resolution, respectively. Candidate sites for preprotein binding are located on a surface containing the SecA epitopes exposed to the periplasm upon binding to SecYEG and are thus positioned to deliver preprotein to SecYEG. Comparisons with structurally related ATPases, including superfamily I and II ATP-dependent helicases, suggest that the interaction geometry of the tandem motor domains in SecA is modulated by nucleotide binding, which is shown by fluorescence anisotropy experiments to reverse an endothermic domain-dissociation reaction hypothesized to gate binding to SecYEG. [ABSTRACT FROM AUTHOR]

Introduction: Morphologic features of the mandible are influenced by the genes of each individual. Mandible size is important to orthodontists because the mandible is the mechanism by which the lower face influences facial esthetics and dental function. To date, no biological marker has been identified that indicates eventual mandible size. This study aimed to correlate the expression of DLX5, DLX6, EDN1, HAND2, PRRX1, and MSX1 to mandible size., Methods: Fifty-nine orthodontic patients aged >6 years who had available cephalometric radiographs were studied. Patients were classified on the basis of condylion-to-gnathion measurements. Messenger RNA was isolated from saliva and subjected to real-time quantitative polymerase chain reaction., Results: Threshold cycle values for subjects with small mandibles (>1 standard deviation [SD] from the mean) had the least expression of DLX6 and MSX1. Threshold cycle values for subjects with large mandibles (>1 SD) had less expression of DLX6 and MSX1 than subjects within 1 SD but more than those with small mandibles., Conclusions: DLX6 and MSX1 are related to mandible development and size. This finding could be used to improve treatment planning for medical and dental professionals seeking to understand the impact of genetics on bone growth., (Copyright © 2022 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Förster resonance energy transfer (FRET) is an established fluorescence-based method used to successfully measure distances in and between biomolecules in vitro as well as within cells. In FRET, the efficiency of energy transfer, measured by changes in fluorescence intensity or lifetime, relates to the distance between two fluorescent molecules or labels. Determination of dynamics and conformational changes from the distances are just some examples of applications of this method to biological systems. Under certain conditions, this methodology can add to and enhance existing X-ray crystal structures by providing information regarding dynamics, flexibility, and adaptation to binding surfaces. We describe the use of FRET and associated distance determinations to elucidate structural properties, through the identification of a binding site or the orientations of dimer subunits. Through judicious choice of labeling sites, and often employment of multiple labeling strategies, we have successfully applied these mapping methods to determine global structural properties in a protein-DNA complex and the SecA-SecYEG protein translocation system. In the SecA-SecYEG system, we have used FRET mapping methods to identify the preprotein-binding site and determine the local conformation of the bound signal sequence region. This study outlines the steps for performing FRET mapping studies, including identification of appropriate labeling sites, discussion of possible labels including non-native amino acid residues, labeling procedures, how to perform measurements, and interpreting the data.

Objectives: To determine three-dimensional treatment changes produced by the Class II Carriere Motion appliance (CMA) in Class II adolescent patients with Class I and Class II skeletal relationships., Materials and Methods: The sample included 59 adolescents (16 boys and 43 girls) with unilateral or bilateral Class II molar and bilateral Class II canine relationship. They were divided into group 1 with skeletal Class I (N = 27; ANB 2.90° ± 1.40°; 13.30 ± 1.53 years) and group 2 with skeletal Class II (N = 32; ANB 6.06° ± 1.64°; 13.26 ± 1.76 years). Cone beam computed tomography images were traced with Invivo software pretreatment (T1) and post-CMA usage (T2). The treatment changes in 36 measurements were calculated in each group, and the changes in 16 measurements were compared between them., Results: In group 1 and 2, maxillary first molars underwent significant distal movement (1.92 mm ± 0.80 mm and 1.67 mm ± 1.56 mm, respectively) with distal tipping and rotation, maxillary canines underwent significant distal movement (2.34 mm ± 1.07 mm and 2.24 mm ± 1.91 mm, respectively) with distal tipping and rotation, and mandibular molars underwent significant mesial movement (-1.37 mm ± 1.23 mm and -2.51 mm ± 1.51 mm, respectively) with mesial tipping. Between the groups, there were significant differences in mandibular molar mesial movement and the U1-SN changes (P < .05)., Conclusions: The CMA corrected Class II malocclusion through distal tipping and rotational movement of maxillary canines and molars and corrected mesial tipping of mandibular molars. Significantly more mandibular molar mesial movement and maxillary incisor flaring were observed in patients with skeletal Class II., (© 2020 by The EH Angle Education and Research Foundation, Inc.)

Background: This study aimed to analyze 3-dimensional nasolabial soft tissue changes following double jaw surgery with Le Fort I maxillary advancement and relate them to underlying surgical movements., Methods: Pre- and post-surgical cone-beam computed tomography images of 37 maxillomandibular surgery patients with ≥2 mm of advancement at A-point were analyzed. Lateral cephalograms were constructed to determine horizontal and vertical surgical movements and 3-dimensional facial images were created from the CBCTs to calculate linear and angular changes., Results: Alar base width increased 1.9 mm, nasal tip protrusion decreased 0.74 mm, the upper lip flattened 7.71 degree and increased 0.9 mm in length, nasal tip was displaced 2.47 mm superiorly causing an increased concavity of 5.52 degree for the overall dorsal surface and 3.99 degree at the supratip break angle (SBA), nasal tip angle decreased 2.16 degree and nasolabial angle decreased 7.37 degree. Correlations were found between underlying surgical movements and nasal tip protrusion, SBA, nasolabial angle (NLA), and nasal tip elevation (NTE). Multiple linear regression equations were calculated to predict changes in SBA from horizontal change at A-point, NLA from horizontal change at B-point, and NTE from the change at A-point horizontally and U1-tip vertically., Conclusion: Double jaw surgery with maxillary advancement has significant effects on nasolabial anatomy.

Objective: The purpose of this study was to assess the long-term stability of rapid palatal expansion (RPE) followed by full fixed edgewise appliances., Methods: This study included 67 patients treated using Haas-type RPE and non-extraction edgewise appliance therapy at a single orthodontic practice. Serial dental casts were obtained at three different time points: pretreatment (T 1 ), after expansion and fixed appliance therapy (T 2 ), and at long-term recall (T 3 ). The mean duration of the T 1 -T 2 and T 2 -T 3 periods was 4.8 ± 3.5 years and 11.0 ± 5.4 years, respectively. The dental casts were digitized, and the computed measurements were compared with untreated reference data., Results: The majority of treatment-related increases in the maxillary and mandibular arch measurements were statistically significant ( p < 0.05) and greater than expected for the untreated controls. Although many measurements decreased postretention (T 2 -T 3 ), the net gains persisted for all of the measurements evaluated., Conclusions: The use of RPE therapy followed by full fixed edgewise appliances is an effective method for increasing maxillary and mandibular arch width dimensions in growing patients., Competing Interests: CONFLICTS OF INTEREST: The authors report no commercial, proprietary, or financial interest in the products or companies described in this article.

Characterization of Sec-dependent bacterial protein transport has often relied on an in vitro protein translocation system comprised in part of Escherichia coli inverted inner membrane vesicles or, more recently, purified SecYEG translocons reconstituted into liposomes using mostly a single substrate (proOmpA). A paper published in this issue (P. Bariya and L. Randall, J Bacteriol 201:e00493-18, 2019, https://doi.org/10.1128/JB.00493-18) finds that inclusion of SecA protein during SecYEG proteoliposome reconstitution dramatically improves the number of active translocons. This experimentally useful and intriguing result that may arise from SecA membrane integration properties is discussed here. Furthermore, determination of the rate-limiting transport step for nine different substrates implicates the mature region distal to the signal peptide in the observed rate constant differences, indicating that more nuanced transport models that respond to differences in protein sequence and structure are needed., (Copyright © 2018 American Society for Microbiology.)

Objectives: To estimate the prevalence of mandibular asymmetries in orthodontic and orthognathic surgery patients and to investigate demographic and skeletal factors associated with this disharmony., Materials and Methods: Cone-beam computed tomography images of 1178 individuals aged 19 through 60 years with complete dentitions were analyzed. Outcomes were classified as relative mandibular symmetry, moderate asymmetry, and severe asymmetry. Factors recorded included sex, age, side of mandibular deviation, sagittal jaw relationship, vertical skeletal pattern, angle of the cranial base, and maxillary asymmetry. Ordinal logistic regression was used to estimate simple and adjusted odds ratios (OR) for the individuals with moderate and severe mandibular asymmetry, as well as 95% confidence intervals., Results: Prevalence values of 55.2%, 27.2%, and 17.6% were observed for relative mandibular symmetry, moderate asymmetry, and severe asymmetry, respectively. An independent association with the side of mandibular deviation and the presence of maxillary asymmetry was observed, both for subjects with moderate mandibular asymmetry (left side: OR = 1.50; 95% CI: 1.01-2.24 / maxillary asymmetry: OR = 2.07; 95% CI: 1.11-3.76) and for those with severe asymmetry (left side: OR = 2.09; 95% CI: 1.27-3.44 / maxillary asymmetry: OR = 4.93; 95% CI: 2.64-9.20)., Conclusions: Moderate and severe mandibular asymmetries were present in 44.8% of the sample, being associated with the side of mandibular deviation and with maxillary asymmetry.

Introduction: The purpose of this study was to determine the accuracy of specific tooth movements with Invisalign (Align Technology, Santa Clara, Calif)., Methods: The study sample included 20 Class I adult patients treated with Invisalign; they completed their first series of aligners and had to have a "refinement" series. Initial and predicted models were obtained from the initial ClinCheck (Align Technology). The starting point of the refinement ClinCheck was used to create the achieved models. Predicted and achieved models were superimposed over the initial ones on posterior teeth using the 3-dimensional Image Analysis open-source software Slicer CMF. Three hundred ninety-eight teeth were measured for vertical, horizontal, and rotational movements, and transverse widths were measured. The amount of predicted tooth movement was compared with the achieved amount for each movement., Results: Horizontal movements of all incisors seemed to be accurate, with small (0.20-0.25 mm) or insignificant differences between predicted and achieved amounts. Vertical movements and particularly intrusions of maxillary central incisors were found to be less accurate, with a median difference of 1.5 mm (P <0.001). All achieved rotations were significantly smaller than those predicted, with the maxillary canines exhibiting the greatest difference of 3.05° (P <0.001)., Conclusions: The most inaccurate movements identified in this study were intrusion of the incisors and rotation of the canines., (Copyright © 2018 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Dens invaginatus is an anomaly of dental development in which calcified tissues, such as enamel and dentin, are invaginated into the pulp cavity. This morphologic alteration is more frequent in maxillary permanent lateral incisors and makes them more susceptible to carious lesions and pulp alterations., Methods: This case report describes a patient with maxillary lateral incisors affected by dens invaginatus. The maxillary right lateral incisor had already undergone endodontic treatment, and the maxillary left one had a periapical lesion. Additionally, the patient had a Class II Division 1 malocclusion, with anterior open bite, posterior crossbite, and an impacted mandibular left second molar., Results: The orthodontic treatment involved extraction of the maxillary lateral incisors and 2 mandibular premolars, resulting in proper overjet and overbite with good arch coordination and occlusal stability., Conclusions: Treatment results were stable, as evaluated in a 6-year posttreatment follow-up., (Copyright © 2018 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: In this study, we investigated volumetric and dimensional changes to the pharyngeal airway space after isolated mandibular setback surgery for patients with Class III skeletal dysplasia., Methods: Records of 28 patients who had undergone combined orthodontic and mandibular setback surgery were obtained. The sample comprised 17 men and 11 women. Their mean age was 23.88 ± 6.57 years (range, 18-52 years). Cone-beam computed tomography scans were obtained at 3 time points: before surgery, average of 6 months after surgery, and average of 1 year after surgery. Oropharyngeal, hypopharyngeal, and total volumes were calculated. The lateral surface and anteroposterior dimensions at the minimal axial areas for oropharyngeal and hypopharyngeal volumes and mean mandibular setback were determined., Results: The mean mandibular setback was 9.93 ± 5.26 mm. Repeated measures analysis of variance determined an overall significant decrease between the means for 6 months and up to 1 year after surgery for oropharyngeal and hypopharyngeal volumes, anteroposterior at oropharyngeal, lateral surface at oropharyngeal, and anteroposterior at hypopharyngeal. No strong correlation between mandibular setback surgery and pharyngeal airway volumes or dimensions was determined., Conclusions: After mandibular setback surgery, pharyngeal airway volume, and transverse and anteroposterior dimensions were decreased. Patients undergoing mandibular setback surgery should be evaluated for obstructive sleep apnea and the proposed treatment plan modified according to the risk for potential airway compromise., (Copyright © 2017 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

A conserved hairpin-like structure comprised of a signal peptide and early mature region initiates protein transport across the SecY or Sec61α channel in Bacteria or Archaea and Eukarya, respectively. When and how this initiator substrate hairpin forms remains a mystery. Here, we have used the bacterial SecA ATPase motor protein and SecYEG channel complex to address this question. Engineering of a functional miniprotein substrate onto the end of SecA allowed us to efficiently form ternary complexes with SecYEG for spectroscopic studies. Förster resonance energy transfer mapping of key residues within this ternary complex demonstrates that the protein substrate adopts a hairpin-like structure immediately adjacent to the SecA two-helix finger subdomain before channel entry. Comparison of ADP and ATP-γS-bound states shows that the signal peptide partially inserts into the SecY channel in the latter state. Our study defines a unique preinsertion intermediate state where the SecA two-helix finger appears to play a role in both templating the substrate hairpin at the channel entrance and promoting its subsequent ATP-dependent insertion., Competing Interests: The authors declare no conflict of interest.

Introduction: This retrospective study included a sample of 300 randomly selected patients from the archived records of Saint Louis University's graduate orthodontic clinic, St. Louis, Mo, from 1990 to 2012. The objective of this study was to quantify the changes obtained in phase 1 of orthodontic treatment and determine how much improvement, if any, has occurred before the initiation of the second phase., Methods: For the purpose of this study, prephase 1 and prephase 2 records of 300 subjects were gathered. All were measured using the American Board of Ortodontics Discrepancy Index (DI), and a score was given for each phase. The difference of the 2 scores indicated the quantitative change of the complexity of the treatment. Paired t tests were used to compare the scores. Additionally, the sample was categorized into 3 groups according to the Angle classifications, and the same statistics were used to identify significant changes between the 2 scores. Analysis of variance was applied to compare the 3 groups and determine which had the most change. Percentages of change were calculated for the significant scores., Results: The total DI score overall and the scores of all 3 groups were significantly reduced from before to after phase 1. Overall, 42% improvement was observed. The Class I group showed 49.3% improvement, the Class II group 34.5% and the Class III group 58.5%. Most components of the DI improved significantly, but a few showed negative changes., Conclusions: Significant reductions of DI scores were observed in the total sample and in all Angle classification groups. This indicates that early treatment reduces the complexity of the malocclusions. Only 2 components of the DI showed statistically significant negative changes., (Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The purposes of this study were to measure the 3-dimensional parameters of the posed smile and to see whether there are any correlations with vertical cephalometric skeletal measurements., Methods: Pretreatment records from a sample of 110 white girls between the ages of 12 and 18 years were gathered. The measurements of SN-GoGn, anterior facial height, and lower and upper facial height percentages were obtained from tracing lateral cephalograms. Superimposing the repose and the posed smile facial scans allowed for measurements to be obtained showing the movements in the x, y, and z dimensions of the upper and lower lips, the commissures, and the Cupid's bows. Correlations and multiple linear regression analyses were run to check for associations and predictive relationships between the cephalometric skeletal measurements and soft tissue changes., Results: We found significant moderate correlations and weak correlations. Significant multiple regression models were found for intercommissural width, smile index, and lower lip in the y and z dimensions., Conclusions: There were moderate correlations showing that as SN-GoGn and anterior facial height increased, the interlabial gap increased as the smile index decreased. Significant relationships were found between certain hard tissue cephalometric measurements and the width of the smile as well as the movements of the lower lip., (Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)