Biomedical subjects

J Huang

Publications and source records attributed to J Huang.

At least 451 records · Page 25Linked to original sources

PubMed1998

Airway hyperreactivity elicited by toluene diisocyanate (TDI)-albumin conjugate is not accompanied by airway eosinophilic infiltration in guinea pigs.

Nonspecific airway hyperresponsiveness is present in many patients with toluene diisocyanate (TDI)-induced asthma; however, the underlying pathophysiological mechanisms of this hyperresponsiveness remain controversial. In the present study, we used a guinea pig model to investigate the association of TDI-induced airway hyperresponsiveness with eosinophilic airway infiltration, which is widely considered to play a key role in the development of allergen-induced hyperresponsiveness. Guinea pigs were sensitized by i.d. injections of 10 microl TDI on day 1 and day 6. Control animals received saline injections. Two weeks after the second injection, airway reactivity to inhaled methacholine and specific airway resistance (sRaw) was measured before and at several times after inhalation challenge with TDI-GSA (guinea pig serum albumin) conjugates. Eosinophils in the airways were detected using enzyme histochemistry and quantified using computer-assisted image analysis. TDI-specific IgG1 antibodies were found in the blood of TDI-sensitized animals. An immediate increase in sRaw was induced in these animals by TDI-GSA challenge; airway hyperresponsiveness to methacholine was observed at 6 h and 18 h after TDI-GSA challenge. However, TDI-GSA challenge did not result in an elevation of eosinophils in the airways, compared with control animals. The results suggest that the development of TDI-induced airway hyperresponsiveness is not dependent upon eosinophil infiltration in airways.

Albumins↗

PubMed1998

Selection and analysis of rare second-site suppressors of Drosophila RNA polymerase II mutations.

We used a mutagenesis and selection procedure in Drosophila melanogaster to recover rare allele-specific suppressor mutations. More than 11 million flies mutant for one of five recessive-lethal mutations in the two largest subunits of RNA polymerase II were selected for additional mutations that restored viability. Forty-one suppressor mutations were recovered. At least 16 are extragenic, identifying a minimum of three loci, two of which do not map near genes known to encode subunits of RNA polymerase II. At most, 25 are intragenic, 4 reverting the initial altered nucleotide back to wild type. Sequence analysis of interacting mutations in the two largest subunits identified a discrete domain in each subunit. These domains might be contact points for the subunits. Finally, our selections were large enough to allow recovery of multiple independent changes in the same nucleotides yet mutations in other equally likely targets were not recovered. The mutations recovered are not random and might provide insights into possible mechanisms for mutagenesis in eukaryotes.

Alleles↗

PubMed1998

Somatostatin stimulates the brush-tailed possum sphincter of Oddi in vitro and in vivo.

BACKGROUND & AIMS: Somatostatin, a neuropeptide and hormone, is found in the biliary tract of several species. The aim of this study was to map the distribution of somatostatin-like immunoreactive nerve fibers in the extrahepatic biliary tract of the Australian possum and to determine the pharmacological effects of somatostatin 1-14 on sphincter of Oddi activity in vitro and in vivo. METHODS: Tissue was harvested for immunohistochemistry and sphincter of Oddi for circular or longitudinal muscle contractility. In anesthetized possums, sphincter of Oddi motility was measured by manometry, and transsphincteric flow was measured gravimetrically. RESULTS: Somatostatin immunoreactivity was evident in gallbladder ganglia nerve cell bodies and in nerve fibers of the common bile duct and sphincter of Oddi. Somatostatin 1-14 increased circular and longitudinal muscle contraction amplitude 3-4-fold (P < 0.05), but only the longitudinal muscle contraction amplitude was tetrodotoxin sensitive. Somatostatin 1-14 stimulated spontaneous sphincter of Oddi motility in a tetrodotoxin-insensitive manner, increasing basal pressure, contraction frequency, and amplitude 2-4-fold (P < 0.05) and reducing transsphincteric flow to 25% of control (P < 0.0001). CONCLUSIONS: Somatostatin-like immunoreactivity is present in the extrahepatic biliary tree, and somatostatin 1-14 stimulates sphincter of Oddi smooth muscle and nerves. The major action is direct stimulation of sphincter of Oddi circular muscle, which reduces transsphincteric flow.

Animals↗

PubMed1998

Fluid resuscitation, nutritional support, and cholesterol in critically ill postsurgical patients.

STUDY OBJECTIVE: To determine how and why cholesterol concentrations decrease after surgery, and the effect of the administration of nutritional support. DESIGN: Prospective, observational study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: 213 consecutive patients admitted to a surgical intensive care unit after major surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Serum concentrations of cholesterol, serum albumin and total protein. The initial portion of this study demonstrated that serum concentrations of cholesterol, like those of serum albumin and total protein, decreased by approximately 30% immediately following surgery. These changes were directly related to changes in hematocrit and inversely correlated with the volume of perioperative intravenous (IV)fluid, the degree of positive fluid balance, and the estimated blood loss. The study's second phase examined the 19 patients who received at least 10 days of nutritional support. After 1 week of feeding, serum total protein concentrations increased significantly, but did not return to preoperative levels. Serum concentrations of cholesterol, which were markedly decreased prior to nutritional repletion, increased significantly after 9 days of treatment. The changes in serum cholesterol concentration were negatively correlated (r = -0.32) with the daily intake of IV fluid. CONCLUSION: Serum cholesterol concentrations, like those of serum albumin and total protein, are markedly reduced immediately following major abdominal surgery, often to levels reported in malnutrition. Dilution by IV fluid and redistribution into an expanded extracellular fluid space are likely the major causes of these decreases. Serum cholesterol concentrations are therefore not useful in the nutritional assessment of patients in the immediate postoperative period.

Adult↗

PubMed1998

Evidence for the participation of the proteasome and calpain in early phases of muscle cell differentiation.

Objectives were to investigate the role of the proteasome and m-calpain to muscle cell differentiation. Accordingly, we investigated the effects of lactacystin, a proteasome inhibitor, and calpain inhibitor-II (CI-II) on L8 muscle cell differentiation and assessed concentrations of proteasomal and calpain subunit mRNAs during differentiation. L8 myoblasts were induced to differentiate by culturing in mitogen-depleted medium. To assess the importance of the proteasome and calpain to differentiation, we examined effects of lactacystin and CI-II on creatine kinase (CK) activity. In the absence of inhibitor, CK activity was detectable within 48 h of mitogen depletion and myotubes were formed. Addition of lactacystin or CI-II to cultures drastically reduced CK activity and prevented formation of myotubes. Hence, proteasome and calpain are both necessary for differentiation. In order to identify which proteasomal subunits were regulated during differentiation, we examined the concentrations of two 20S core subunits (C8 and C9) and three 22S ATPases (MSS1, S4 and TBP1) during differentiation. Concentrations of m-calpain and beta-tubulin mRNAs were also assessed. Differentiation was associated with slight increases (ca. 30%) in concentrations of mRNAs encoding the proteasomal 20S core subunits (C8 and C9) and with large increases (approximately 2-fold) in mRNAs encoding the regulatory subunit ATPases. m-calpain mRNA concentration also increased two-fold following mitogen depletion. beta-Tubulin mRNA concentration remained unchanged early in the differentiation process and thereafter declined. Of interest, changes in proteasomal and m-calpain mRNAs occurred within 6-24 h of mitogen depletion (i.e., at least 24-36 h prior to detectable changes in creatine kinase activity). These results indicate that changes in expression of proteasome and calpains subunits occur early in the differentiation process. These changes may be required for the normal course of differentiation to proceed. Differentiation is associated with larger changes in proteasomal ATPase mRNAs than in 20S core particle mRNAs indicating that either turnover rates of the 22S ATPase subunits are more rapid in differentiating cells than of the 20S core particles or that functions of the regulatory subunits become more important during muscle cell differentiation.

Acetylcysteine↗

PubMed1998

Protein and mRNA analysis of myosin heavy chains in the developing avian pectoralis major muscle.

While the existence of post-hatch and adult myosin heavy chain isoforms in the large, avian type IIB pectoralis major muscle has been clearly established, the number and nature of fast myosin heavy chains during in ovo development and the perihatch period have not been resolved. In the present study, developmental fast heavy chain proteins purified by high resolution anion-exchange have been characterized by sequence analysis of a unique CNBr peptide and by complementary mRNA analysis. The four proteins present at 15/16 days in ovo are shown to differ uniquely in primary structure. They correlate with heavy chains II, IV, VI and VII, characterized recently as major or minor species in adult fast muscles using similar methods. These four heavy chains are expressed in a time-dependent fashion from 8 to 16 days in ovo. At the mRNA level, heavy chain VI predominates until 12 days in ovo. Heavy chain IV mRNA is upregulated dramatically at 16 days in ovo preparatory to its protein's predominance in the peri-hatch period. Heavy chains II, IV and V (the post-hatch isoform which replaces heavy chain IV) have major roles in adult fast muscles.

Animals↗

PubMed1998

Cell death attenuation by 'Usurpin', a mammalian DED-caspase homologue that precludes caspase-8 recruitment and activation by the CD-95 (Fas, APO-1) receptor complex.

Apoptotic cell suicide initiated by ligation of CD95 (Fas/APO-1) occurs through recruitment, oligomerization and autocatalytic activation of the cysteine protease, caspase-8 (MACH, FLICE, Mch5). An endogenous mammalian regulator of this process, named Usurpin, has been identified (aliases for Usurpin include CASH, Casper, CLARP, FLAME-1, FLIP, I-FLICE and MRIT). This protein is ubiquitously expressed and exists as at least three isoforms arising by alternative mRNA splicing. The Usurpin gene is comprised of 13 exons and is clustered within approximately 200 Kb with the caspase-8 and -10 genes on human chromosome 2q33-34. The Usurpin polypeptide has features in common with pro-caspase-8 and -10, including tandem 'death effector domains' on the N-terminus of a large subunit/small subunit caspase-like domain, but it lacks key residues that are necessary for caspase proteolytic activity, including the His and Cys which form the catalytic substrates diad, and residues that stabilize the P1 aspartic acid in substrates. Retro-mutation of these residues to functional caspase counterparts failed to restore proteolytic activity, indicating that other determinants also ensure the absence of catalytic potential. Usurpin heterodimerized with pro-caspase-8 in vitro and precluded pro-caspase-8 recruitment by the FADD/MORT1 adapter protein. Cell death induced by CD95 (Fas/APO-1) ligation was attenuated in cells transfected with Usurpin. In vivo, a Usurpin deficit was found in cardiac infarcts where TUNEL-positive myocytes and active caspase-3 expression were prominent following ischemia/reperfusion injury. In contrast, abundant Usurpin expression (and a caspase-3 deficit) occurred in surrounding unaffected cardiac tissue, suggesting reciprocal regulation of these pro- and anti-apoptotic molecules in vivo. Usurpin thus appears to be an endogenous modulator of apoptosis sensitivity in mammalian cells, including the susceptibility of cardiac myocytes to apoptotic death following ischemia/ reperfusion injury.

Alternative Splicing↗

PubMed1998

Semiparametric linkage analysis using pseudolikelihoods on neighbouring sets.

For many complex diseases, study has suggested that the disease genes influence not only the occurrence of the disease, but also the age of onset. Current methods in linkage analysis are mainly concentrated on affected relative pairs or affected family members, and age of onset information is either ignored or is taken into account by specifying age-dependent penetrances for liability classes. In fact, affected relatives with different ages of onset may be the result of different genetic aetiologies and unaffected relatives are censored at the study time. Therefore, incorporation of age of onset and including contrasts between affected and unaffected pedigree members are important components of effective analysis for the detection of linkage with genetic markers. We use multiple markers to infer the inheritance vector in order to extract information about the inheritance pattern of the disease allele in a pedigree. For a given inheritance vector, we define two neighbour sets for each individual based on allele identical by descent (IBD). We then use the within-set and between-sets conditional hazard ratios to characterize the dependence of age of onset among relatives. A pseudolikelihood ratio test is proposed for testing linkage. Simulated and real data sets are used to illustrate these new statistical methods.

Age of Onset↗

PubMed1998

Lupeol is the cytotoxic principle in the leaf extract of Dendropanax cf. querceti.

The crude ethanol extract from the leaves of Dendropanax cf. querceti (Araliaceae) from Monteverde, Costa Rica, exhibits cytotoxic activity against Hep-G2, A-431, and H-4IIE tumor cell lines. The active component has been isolated by activity-directed separation and identified by 1H- and 13C-NMR spectroscopy as the triterpene lupeol. The mechanism of cytotoxic activity of lupeol has been determined to be inhibition of topoisomerase II.

Animals↗

PubMed1998

Operative management of islet-cell tumors arising in the head of the pancreas.

BACKGROUND: Pancreatic islet cell tumors (ICTs) can be treated by enucleation or pancreatic resection. We reviewed our experience with ICTs in the head of the pancreas to define indications for enucleation versus pancreaticoduodenectomy. METHODS: Between January 1982 and December 1997, 48 patients underwent surgical resection for presumed ICTs of the pancreatic head. Of these, 18 were found on pathologic examination to be disease in a lymph node. Thirty patients had 32 true pancreatic head ICTs. We reviewed the operative results and postoperative courses in these patients. RESULTS: Mean diameter of the ICTs was 15 mm. Twenty-seven patients (90%) underwent successful enucleations. Three patients failed enucleation and underwent pancreaticoduodenectomy. There was no operative mortality. The median time to regular diet was 7 days. The median time to removal of all drains was 22 days. The most common complication was pancreatic fistula (15%). No patient required reoperation for treatment of a fistula. CONCLUSION: Most ICTs of the pancreatic head can be removed with enucleation, preserving pancreatic tissue and avoiding the morbidity of pancreaticoduodenectomy. The primary indication for pancreaticoduodenectomy is not the size of the lesion but its proximity to the pancreatic duct.

Adolescent↗

PubMed1998

Statistical evaluation of age-at-onset anticipation: a new test and evaluation of its behavior in realistic applications.

The discovery that microsatellite repeat expansions can cause clinical disease has fostered renewed interest in testing for age-at-onset anticipation (AOA). A commonly used procedure is to sample affected parent-child pairs (APCPs) from available data sets and to test for a difference in mean age at onset between the parents and the children. However, standard statistical methods fail to take into account the right truncation of both the parent and child age-at-onset distributions under this design, with the result that type I error rates can be inflated substantially. Previously, we had introduced a new test, based on the correct, bivariate right-truncated, age-at-onset distribution. We showed that this test has the correct type I error rate for random APCPs, even for quite small samples. However, in that paper, we did not consider two key statistical complications that arise when the test is applied to realistic data. First, affected pairs usually are sampled from pedigrees preferentially selected for the presence of multiple affected individuals. In this paper, we show that this will tend to inflate the type I error rate of the test. Second, we consider the appropriate probability model under the alternative hypothesis of true AOA due to an expanding microsatellite mechanism, and we show that there is good reason to believe that the power to detect AOA may be quite small, even for substantial effect sizes. When the type I error rate of the test is high relative to the power, interpretation of test results becomes problematic. We conclude that, in many applications, AOA tests based on APCPs may not yield meaningful results.

Age Factors↗

PubMed1998

Pharmacokinetics and pharmacodynamics of SM-20302, a GPIIb/IIIa receptor antagonist, in anesthetized dogs.

We examined the pharmacokinetic and pharmacodynamic properties of SM-20302, a GPIIb/IIIa receptor antagonist, in anesthetized dogs. SM-20302 was administered intravenously in doses of 30 (n=2), 100 (n=4), 300 (n=4), and 1,000 microg/kg (n=4). The half-life of the initial phase was 4 min, and that of the terminal phase was 162-209 min. SM-20302 produced a dose-dependent increase in the initial plasma concentration and the area under concentration-time curve but did not alter the volume of distribution, mean residence time, or plasma clearance. Plasma clearance for SM-20302 ranged from 6.58 to 9.73 ml/min/kg. All doses of SM-20302 inhibited (> or =90%) the ex vivo platelet aggregation induced by adenosine diphosphate (ADP) or arachidonic acid (AA) in citrated platelet-rich plasma (cPRP). In heparinized PRP (hPRP), a dose-dependent (44-89%) inhibition was observed. By using a sigmoid Emax model, the in vivo median inhibitory concentration (IC50) for SM-20302 was estimated to be 14-19 ng/ml in cPRP and 79-89 ng/ml in hPRP. To validate the calculated parameters, an infusion regimen was designed for the prevention of coronary artery thrombosis. Infusion of SM-20302 produced 64-67% inhibition of platelets in hPRP and maintained vessel patency despite vessel wall injury. The results suggest that SM-20302 exhibits linear pharmacokinetics and that its ability to inhibit platelet aggregation in hPRP may correlate more accurately with its in vivo antithrombotic efficacy.

Animals↗

PubMed1998

Evolution of the organization of epicardial activation patterns during ventricular fibrillation.

INTRODUCTION: This study quantified how the organization of epicardial activation changes during the first 40 seconds of ventricular fibrillation (VF). METHODS AND RESULTS: Unipolar potentials were mapped from a 504 (24 x 21) electrode array (2-mm interelectrode spacing) on the anterior right ventricle (RV) and left ventricle (LV) epicardium. The array covered approximately 20% of the epicardial surface. In each of seven pigs, six episodes of VF were induced by premature stimulation. One-half second epochs of VF were analyzed, starting 0, 10, 20, 30, and 40 seconds post induction and using novel pattern analysis algorithms. Eight parameters were quantified: (1) the number of wavefronts; (2) the epicardial area activated by wavefronts; (3) the fraction of wavefronts arising from epicardial breakthrough or from a focus; (4) the fraction of wavefronts terminated by conduction block; (5) the multiplicity index (number of distinct activation pathways in the rhythm); (6) the repeatability index (number of times activation pathways are traversed); (7) the activation rate; and (8) the wavefront propagation velocity. The results showed that VF patterns were less organized at 10 than at 0 seconds, with more, smaller wavefronts traversing a larger variety of pathways for fewer repetitions. VF activation patterns then gradually reorganized up to 40 seconds, but by a different mechanism: the spatial size of subpatterns grew, but the dynamics otherwise appeared unchanged. During both transitions, both activation rate and propagation velocity slowed monotonically. CONCLUSION: Thus, changes in organization during VF can occur by multiple mechanisms.

Action Potentials↗

PubMed1998

Methamphetamine-induced modification of dopamine metabolism in cultured striatal astrocytes.

The role of striatal astrocytes in the metabolic processing (by deamination) of methamphetamine-released dopamine is not known. To investigate the relationship between methamphetamine and dopamine metabolism, we measured 6-hydroxydopamine, dopamine and, 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations following methamphetamine treatment of cultured striatal astrocytes prepared from 1-2 day-old rats. Addition of low concentrations of dopamine (5 x 10(-5) to 5 x 10(-4) M) to cultured astrocytes increased DOPAC levels in a dose-dependent fashion while higher concentrations (5 x 10(-3) to 10(-2) M) inhibited its metabolism and induced formation of 6-hydroxydopamine. Under the same experimental conditions, 10(-4) M dopamine in combination with methamphetamine (10(-5) to 10(-3) M) inhibited DOPAC formation and increased dopamine levels in a dose-dependent fashion, but the formation of intracellular 6-hydroxydopamine was not evident. Deprenyl (10(-5) or 10(-4) M), an inhibitor of monoamine oxidase B, and pargyline (10(-5) or 10(-4) M), a non-selective monoamine oxidase inhibitor, completely inhibited DOPAC formation and increased dopamine levels, while clorgyline (10(-5) or 10(-4) M), an inhibitor of monoamine oxidase-A, only partially inhibited DOPAC formation (42 or 45% of control, respectively). These results support the hypothesis that methamphetamine inhibits monoamine oxidase and causes increases in dopamine levels in cultured striatal astrocytes.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1998

Mannose induces the release of cytopathic factors from Acanthamoeba castellanii.

Acanthamoeba keratitis is a chronic inflammatory disease of the cornea which is highly resistant to many antimicrobial agents. The pathogenic mechanisms of this disease are poorly understood. However, it is believed that the initial phases in the pathogenesis of Acanthamoeba keratitis involve parasite binding and lysis of the corneal epithelium. These processes were examined in vitro, using Acanthamoeba castellanii trophozoites. Parasites readily adhered to Chinese hamster corneal epithelial cells in vitro; however, parasite binding was strongly inhibited by mannose but not by lactose. Although mannose prevented trophozoite binding, it did not affect cytolysis of corneal epithelial cells. Moreover, mannose treatment induced trophozoites to release cytolytic factors that lysed corneal epithelial cells in vitro. These factors were uniquely induced by mannose because supernatants collected from either untreated trophozoites or trophozoites treated with other sugars failed to lyse corneal cells. The soluble factors were size fractionated in centrifugal concentrators and found to be > or = 100 kDa. Treatment of the supernatants with the serine protease inhibitor phenylmethylsulfonyl fluoride inhibited most, but not all, of the cytopathic activity. These data suggest that the binding of Acanthamoeba to mannosylated proteins on the corneal epithelium may exacerbate the pathogenic cascade by initiating the release of cytolytic factors.

Acanthamoeba↗

PubMed1998

Joint transcriptional control of xpsR, the unusual signal integrator of the Ralstonia solanacearum virulence gene regulatory network, by a response regulator and a LysR-type transcriptional activator.

Ralstonia (Pseudomonas) solanacearum is a soil-borne phytopathogen that causes a wilting disease of many important crops. It makes large amounts of the exopolysaccharide EPS I, which it requires for efficient colonization, wilting, and killing of plants. Transcription of the eps operon, encoding biosynthetic enzymes for EPS I, is controlled by a unique and complex sensory network that responds to multiple environmental signals. This network is comprised of the novel transcriptional activator XpsR, three distinct two-component regulatory systems (VsrAD, VsrBC, and PhcSR), and the LysR-type regulator PhcA, which is under the control of PhcSR. Here we show that the xpsR promoter (PxpsR) is simultaneously controlled by PhcA and VsrD, permitting XpsR to act like a signal integrator, simultaneously coordinating signal input into the eps promoter from both VsrAD and PhcSR. Additionally, we used in vivo expression analysis and in vitro DNA binding assays with substitution and deletion mutants of PxpsR to show the following. (i) PhcA primarily interacts with a typical 14-bp LysR-type consensus sequence around position -77, causing a sixfold activation of PxpsR; a weaker, less-defined binding site between -183 and -239 likely enhances PhcA binding and activation via the -77 site another twofold. (ii) Full 70-fold activation of PxpsR requires the additional interaction of the VsrD response regulator (or its surrogate) with a 14-bp dyadic sequence centered around -315 where it enhances activation (and possibly binding) by PhcA; however, VsrD alone cannot activate PxpsR. (iii) Increasing the distance between the putative VsrD binding site from that of PhcA by up to 232 bp did not dramatically affect PxpsR activation or regulation.

Bacterial Proteins↗

PubMed1998

Comparisons of highly virulent H5N1 influenza A viruses isolated from humans and chickens from Hong Kong.

Genes of an influenza A (H5N1) virus from a human in Hong Kong isolated in May 1997 were sequenced and found to be all avian-like (K. Subbarao et al., Science 279:393-395, 1998). Gene sequences of this human isolate were compared to those of a highly pathogenic chicken H5N1 influenza virus isolated from Hong Kong in April 1997. Sequence comparisons of all eight RNA segments from the two viruses show greater than 99% sequence identity between them. However, neither isolate's gene sequence was closely (>95% sequence identity) related to any other gene sequences found in the GenBank database. Phylogenetic analysis demonstrated that the nucleotide sequences of at least four of the eight RNA segments clustered with Eurasian origin avian influenza viruses. The hemagglutinin gene phylogenetic analysis also included the sequences from an additional three human and two chicken H5N1 virus isolates from Hong Kong, and the isolates separated into two closely related groups. However, no single amino acid change separated the chicken origin and human origin isolates, but they all contained multiple basic amino acids at the hemagglutinin cleavage site, which is associated with a highly pathogenic phenotype in poultry. In experimental intravenous inoculation studies with chickens, all seven viruses were highly pathogenic, killing most birds within 24 h. All infected chickens had virtually identical pathologic lesions, including moderate to severe diffuse edema and interstitial pneumonitis. Viral nucleoprotein was most frequently demonstrated in vascular endothelium, macrophages, heterophils, and cardiac myocytes. Asphyxiation from pulmonary edema and generalized cardiovascular collapse were the most likely pathogenic mechanisms responsible for illness and death. In summary, a small number of changes in hemagglutinin gene sequences defined two closely related subgroups, with both subgroups having human and chicken members, among the seven viruses examined from Hong Kong, and all seven viruses were highly pathogenic in chickens and caused similar lesions in experimental inoculations.

Amino Acid Sequence↗

PubMed1998

Intramolecular regulation of MyoD activation domain conformation and function.

The MyoD family of basic helix-loop-helix (bHLH) proteins is required for myogenic determination and differentiation. The basic region carries the myogenic code and DNA binding specificity, while the N terminus contains a potent transcriptional activation domain. Myogenic activation is abolished when the basic region, bound to a myogenic E box, carries a mutation of Ala-114. It has been proposed that DNA binding of the MyoD basic region leads to recruitment of a recognition factor that unmasks the activation domain. Here we demonstrate that an A114N mutant exhibits an altered conformation in the basic region and that this local conformational difference can lead to a more global change affecting the conformation of the activation domain. This suggests that the deleterious effects of this class of mutations may result directly from defective conformation. Thus, the activation domain is unmasked only upon DNA binding by the correct basic region. Such a coupled conformational relationship may have evolved to restrict myogenic specificity to a small number of bHLH proteins among many with diverse functions yet with DNA binding specificities known to be similar.

Alanine↗