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PubMed1993

Rab11, a small GTPase associated with both constitutive and regulated secretory pathways in PC12 cells.

A specific polyclonal antibody was used to investigate the subcellular distribution of the small GTPase, rab11p, in the neuroendocrine cell line, PC12. We took advantage of a previously described pulse-chase protocol based on sulfation to examine the distribution of rab11 along the secretory pathway. Using the rab11 antiserum, but not serum depleted of rab11 antibodies, we were able to specifically immunoisolate markers of the constitutive and the regulated secretory pathway in the trans-Golgi network (TGN) as well as after their exit from this compartment (constitutive secretory vesicles, immature, and mature secretory granules). We therefore conclude that rab11p is associated with the TGN and with TGN-derived vesicles of both the constitutive and the regulated secretory pathway in PC12 cells.

Animals↗

PubMed1996

Differential effects of NaCl concentration on the constitutive activity of the thyrotropin and the luteinizing hormone/chorionic gonadotropin receptors.

The TSH receptor (TSHR) and the LH/CG receptor (LHR) are members of the family of G protein-coupled receptors. Recently, point mutations conferring constitutive activity to the TSHR and LHR have been observed as a cause of toxic adenoma and familial/sporadic male pseudo-precocious puberty, respectively. When evaluated by transfection in COS-7 cells the wild-type (wt) TSHR displays definite constitutive activity towards Gs-dependent adenylylcyclase stimulation, while available evidence shows that the LHR does not. In order to compare the constitutive activity of both receptors, we performed functional studies in COS-7 cells using different assay conditions. Human TSHR and LHR cDNAs subcloned in the expression vector pSVL were transiently expressed in COS-7 cells and cAMP production was determined following incubation in a medium containing physiological concentration of NaCl [isotonic (NaCl)] or in the same medium without NaCl [hypotonic (NaCl-)] or where NaCl was replaced by an isoosmolar concentration of sucrose [isotonic (sucrose)]. Cells transfected with the TSHR showed higher basal cAMP levels over cells transfected with pSVL in all conditions tested. The effect was stronger when cells were incubated in isotonic (sucrose) buffer. Cells expressing LHR exhibited a minimal increase of cAMP levels over cells transfected with pSVL in isotonic (NaCl) buffer; however, a marked increase in basal cAMP levels was observed when cells were assayed in hypotonic (NaCl-) or isotonic (sucrose) buffers. Varying the pH or incubation temperature was without effect on the results obtained with both receptors. Our data show that despite extensive sequence similarity, the LH and TSH receptors differ markedly in their basal activity. The differential sensitivity of both receptors to low NaCl concentrations, suggests that the unliganded TSH receptor is less constrained than its LH homolog and may be more susceptible to activation by a wide spectrum of mutations.

Animals↗

PubMed1996

Prostaglandin E receptor EP3gamma isoform, with mostly full constitutive Gi activity and agonist-dependent Gs activity.

We recently demonstrated that two exclusively Gi-coupled isoforms of the mouse EP3 receptor, EP3alpha and beta, with different carboxyl-terminal tails, differed in agonist-independent constitutive Gi activity, and the carboxyl-terminal tail-truncated receptor showed full constitutive activity (Hasegawa, H., Negishi, M., and Ichikawa, A. (1996) J. Biol. Chem. 271, 1857-1860). Here we further examined Gi and Gs activities of the third isoform, EP3gamma, coupled to both Gi and Gs. The My receptor showed mostly full constitutive Gi activity and agonist-dependent Gs activity. The truncated receptor also showed agonist-dependent Gs activity, but the level was lower than that of the EP3gamma receptor. Thus, the carboxyl-terminal tail would differentially regulate Gi and Gs activities of the EP3 receptor.

Adenylyl Cyclases↗

PubMed1985

Constitutive stress--strain relations for the myocardium in diastole.

The importance of stress-strain myocardial constitutive relations is that they provide a criterion for behavior in vivo. Our purpose was to develop constitutive equations which are valid in diastole. The myocardium was assumed to be composed of a nonlinear viscoelastic, inhomogeneous, anisotropic (transversely isotropic) and incompressible material operating under adiabatic and isothermal conditions. The expressions contain five moduli. Two are fixed by the restriction of incompressibility, one is estimated, the remaining two refer to directions along and perpendicular to a fiber. Both possess a bimodal variation with intermodal switching occurring in late rapid filling and diastasis. They are functions of time and material constants. These constants can be observed. A dynamic test is suggested. Constitutive statements complete a set of equations sufficient for the solution of a class of boundary value problems. One type is formulated. They also permit the determination of stress from measured strain. Examples are given.

Heart↗

PubMed1986

A constitutive model for two-dimensional soft tissues and its application to experimental data.

A constitutive relation proposed by Shoemaker (Ph.D. dissertation, 1984) to model the mechanical behavior of membraneous or two-dimensional soft tissues is described. Experiments by Schneider (Ph.D. dissertation, 1982) on human skin and Lee et al. (Am. J. Physiol., 249, H222-H230, 1985) on canine pericardium, and the application of the constitutive model to biaxial stress-strain data from these experiments, are discussed. Some experimental data and predictions of the model obtained by curvefitting are presented for comparison. Values of material parameters are also presented. It is concluded that the constitutive model is well able to fit results of individual tests, and that its generality (judged by consistency of parameters from test to test of the same specimen), though not complete, does compare favorably with some other results presented in the literature.

Animals↗

PubMed1990

Cell-free protein sorting to the regulated and constitutive secretory pathways.

To elucidate the mechanism of secretory granule formation, we here identify the first intermediate in this process, the immature secretory granule, in the neuroendocrine cell line PC12 and demonstrate the packaging of a regulated secretory protein, secretogranin II, to immature secretory granules in a cell-free system. The formation of immature secretory granules was as fast (t1/2 approximately 5 min) as that of constitutive secretory vesicles identified by the presence of a rapidly secreted heparan sulfate proteoglycan. Using the cell-free system, the formation of post-Golgi secretory vesicles was found to be dependent upon ATP. Two distinct populations of vesicles were formed: immature secretory granules containing secretogranin II and constitutive secretory vesicles containing the heparan sulfate proteoglycan. These results show that in a cell-free system, a constitutive and a regulated secretory protein are sorted upon exit from the trans-Golgi network.

Adenosine Triphosphate↗

PubMed1982

Abnormalities of chromosome #13 in retinoblastomas from individuals with normal constitutional karyotypes.

Constitutional chromosome abnormalities have been associated with retinoblastoma, Wilm's tumor, and a familial form of renal carcinoma. For each tumor type, the particular chromosome segment involved in the observed rearrangements is different: in retinoblastoma, that segment is band q14 on chromosome #13. We now present evidence that in retinoblastoma, structural abnormalities involving the particular chromosome segment identified in the constitutional cases can also occur in the tumors of individuals with normal constitutional karyotypes. Six cases with retinoblastoma in one or both eyes were analyzed; deletions/rearrangements involving 13q14 were found in the tumor cell karyotypes of five of the six. These observations suggest that changes in a gene or genes at a common site (13q14) play a role in tumorigenesis in all forms of retinoblastoma, sporadic as well as heritable.

Child, Preschool↗

PubMed1985

Cytogenetic pattern in leukemic cells of patients with constitutional chromosome anomalies.

Acquired karyotypic changes analyzed by banding techniques in 21 patients with a malignant hematologic disorder and a major constitutional chromosome anomaly, including ten patients with trisomy 21, five patients with a balanced translocation, and six patients with a sex chromosome anomaly. Detailed karyotypic findings were ascertained in 28 additional patients reported in the literature. Some striking differences were observed in the combined material of the present series and cases previously published as regards (a) distribution of morphological leukemia types among patients with different types of constitutional anomalies, and (b) incidence and type of acquired chromosomal abnormality among patients with different types of constitutional anomalies.

Adolescent↗

PubMed1987

Frequency of constitutional chromosome alterations in patients with hematologic neoplasias.

From 1978 to 1985 cytogenetic studies were performed on 718 patients with different hematologic diseases. Nine (1.25%) had a constitutional chromosome alteration. One patient had trisomy 21, four had balanced translocations and four had sex chromosome anomalies. Although the frequency of constitutional alterations was twice that seen in the newborn population, an analysis of these data and also from the literature shows a random association between constitutional chromosome alterations and hematologic neoplasias, except for patients with Down's syndrome.

Adolescent↗

PubMed1989

Acute nonlymphocytic leukemia in a patient with a constitutional inv(4).

We describe a case of acute nonlymphocytic leukemia (ANLL) in a patient with a constitutional chromosome anomaly, inv(4)(p16q26). The patient had extensive occupational exposure to toxic chemicals. Reports of constitutional or acquired chromosome inversions in human malignancies are quite uncommon. The constitutional changes associated with hematologic malignancies include trisomy 21, balanced translocations, deletions, and sex chromosome anomalies. The breakpoints on chromosome 4 in our case are 4p16, to which the murine leukemia viral (v-raf) oncogene, pseudogene 1, has been mapped, and 4q26, which is the locus of the IL-2 gene. Activation of these genes could have played a role in the pathogenesis of the patient's leukemia.

Aged↗

PubMed1995

No statistical association between fragile sites and constitutional chromosome breakpoints.

Ten thousand four hundred ninety-two constitutional breakpoints available from the cytogenetic literature were analyzed for their coincidence with known fragile sites (FS) at 303-band resolution. In this analysis we have taken into account the stochastic connections of some features of chromosome bands with both the presence of FS and constitutional breakage. Our results suggest that there is no particular association between FS and constitutional chromosome rearrangements.

Chromosome Aberrations↗

PubMed1995

Regulation of the inducible soybean nitrate reductase isoform in mutants lacking constitutive isoform(s).

In soybean, three nitrate reductase isoforms have been identified based on metabolic regulation, substrate specificity, and kinetic parameters. Two isoforms have been termed constitutive, as their activities are present without the addition exogenous nitrate to soybean seedlings. The third activity is termed inducible, as its activity is present only when soybean plants have been supplied with nitrate. The purpose of this study was to examine the regulation of the inducible nitrate reductase isoform in soybean mutants lacking one or both of the constitutive isoforms. Based on evidence obtained through measurements of enzyme activity, Western blotting, and RNA determinations, the absence of one or both of the constitutive nitrate reductase isoforms has no effect on the metabolite regulation of the inducible nitrate reductase isoform.

Enzyme Induction↗

PubMed1989

Constitutive c-myc expression enhances proliferation of differentiating F9 teratocarcinoma cells.

The c-myc protooncogene is expressed in many tumor cells as well as during normal development. In order to study the role of c-myc in differentiation, proliferation and tumorigenicity of F9 mouse teratocarcinoma cells, the pSVmyc1 plasmid constitutively expressing an active c-myc oncogene was introduced into F9 stem cells by cotransfection with the selectable marker RSVneo. Enhanced expression of c-myc did not alter the properties of F9 stem cells. Prolonged proliferation during retinoic acid induced differentiation was observed in cell clones constitutively expressing c-myc. In contrast, as determined by morphology, by immunocytochemistry for markers specific for stem cells and differentiated derivatives, and by Northern hybridization for mRNAs specific for differentiated cells, differentiation was neither inhibited nor delayed by constitutive c-myc expression. Tumorigenicity of stem cells as well as retinoic acid-treated cells--as measured by soft agar cloning efficiency and tumor formation in syngenic mice--was not altered by SVmyc1. We conclude that in F9 teratocarcinoma cells down-regulation of c-myc is related to arrest of proliferation rather than differentiation.

Animals↗

PubMed1996

[Renal vein thrombosis and constitutional protein S deficiency].

Venous and arterial thrombosis due to a constitutional protein S deficiency is well-known. We report the case of a 36 year-old patient admitted to hospital in 1991 for primary renal vein thrombosis due to a constitutional protein S deficiency of type I. The diagnosis was made by CT scan and angiography. Left nephrectomy, which was made because of doubt with regard to subjacent neoplasm, showed left renal vein thrombosis and multiple renal infarcts. In 1994, after 4 months of discontinuation of oral anticoagulants, the patient presented pulmonary embolism documented by pulmonary scintigraphy and CT scan, partial portal thrombosis and sural thrombophlebitis documented by echography coupled with Doppler. To our knowledge, this is the first reported case of a constitutional protein S deficiency associated with primary renal vein thrombosis.

Adult↗

PubMed1990

A Staphylococcus aureus plasmid that specifies constitutive macrolide-lincosamide-streptogramin B resistance contains a novel deletion in the ermC attenuator.

A 2.5 kb plasmid, pA22, isolated from a naturally occurring S. aureus strain confers constitutive MLS-resistance. By restriction enzyme analysis, pA22 is indistinguishable from the S. aureus inducible MLS-resistance conferring plasmid, pT48, apart froma small deletion. DNA sequencing showed that the deletion, is in the leader/attenuator region of the ermC (MLS-resistance) gene and removes some of the complementary repeat regions required by the translational attenuation model in pT48 for inducible ermC expression. The deletion in plasmid pA22 is different from that found in similar 2.5kb constitutive MLS-resistance plasmids in other Gram-positive bacteria. It is suggested that plasmids conferring the constitutive phenotype have evolved from an inducible ancestor on several independent occasions.

Amino Acid Sequence↗

PubMed1977

Insertion sequence IS2 associated with int-constitutive mutants of bacteriophage lambda.

We have examined mutations in bacteriophage lambda called int-c, which confer elevated constitutive expression on the int gene for prophage integration. One class of mutations, which map between the b538 and bio386 endpoints, does not appear to be associated with any major chromosomal modification, whereas the second class has the IS2 insertion sequence in orientation II within the region between gene int and the b538 endpoint, All int-c mutations are within gene xis, with the possible exception of int-c548, which might be located between int and xis. The present data are most consistent with the following notion: (1) the point mutations of class one inactivate the tI terminator signal of the pI-tI leader RNA for gene int and thus render int expression independent of the antiterminating action of the cII and cIII products, and (2) the second class of int-c mutants is constitutive for Int because the IS2 insertion, when strategically located between int and tI, provides a new constitutive promoter for int transciption.

Coliphages↗

PubMed1988

Ammonia-constitutive nitrogen fixation mutants of Rhodobacter capsulatus.

Mutants of R. capsulatus that express nif genes constitutively with respect to ammonia were studied in order to define better the circuit that regulates nif gene transcription. One mutant class could be complemented in trans by a cosmid clone containing a wild-type gene (nifR5) defined by Tn5 inserts as being no longer than 1.6 kb. The nifR5 gene is unlinked to previously described nif genes. A second mutant class could not be complemented by the wild-type cosmid library. For one mutant in this class, a nifH::lac fusion was used to select further mutants that were Lac-. Only two of these could be complemented in trans to Lac+; the complementing gene was nifR4, which is analogous to the ntrA gene of enterobacteria. Both complemented strains were Nifc. Therefore these mutations do not bypass the need for the nifR4 gene product. A third class of constitutive mutant was found by selecting Nif+ revertants of a Nif- strain deleted for the nifR1 and nifR2 genes. The nifR1 and nifR2 genes are homologues of enterobacterial ntrC and ntrB genes, respectively. Not all of the Nif+ revertants were constitutive; some were regulated normally by ammonia. We suspect that the latter revertants use alternate Ntr systems to activate nif gene transcription, a suggestion consistent with the observation that numerous bands in Southern blots of total DNA of R. capsulatus are identified by Escherichia coli ntr gene probes.

Ammonia↗

PubMed1990

Isolation of a Bacillus subtilis mutant defective in constitutive O6-alkylguanine-DNA alkyltransferase.

A mutant of Bacillus subtilis defective in the constitutive activity of O6-alkylguanine-DNA alkyltransferase was isolated from a strain (ada-1) deficient in the adaptive response to DNA alkylation. Cells carrying the mutation dat-1 which was responsible for the defect in constitutive activity exhibited hypersensitivity for lethality and mutagenesis when challenged with methyl-nitroso compounds. The constitutive activity is independent of the adaptive response, and seems to function as a basal defense against environmental alkylating agents.

Alkyl and Aryl Transferases↗