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PubMed2005

Rate of chromosome changes and speciation in reptiles.

The chromosome changing rate (i.e. the number of chromosome rearrangements per million years) was studied in 1,329 reptile species in order to evaluate the karyological evolutionary trend and the existence of possible correlations between chromosome mutations and some aspects of the evolution of this class. The results obtained highlight the existence of a general direct correlation between chromosome changing rate and number of living species, although different trends can be observed in the different orders and suborders. In turtles, the separation of pleurodires from cryptodires was accompanied by a considerable karyological diversification. Among pleurodires, the evolution of the Chelidae and Pelomedusidae was also characterised by chromosome variation, while in cryptodires a marked karyological homogeneity is observed between and within infra-orders. Similarly there is no correlation between changing rate and species number in crocodiles, where the evolution of the families and genera has entailed few chromosome mutations. Chromosome variability was greater in lizards and snakes. In the formers variations in chromosome changing rate accompanied the separation of the infra-orders and the evolution of most of the families and of some genera. The origin of snakes has also been accompanied by a marked karyological diversification, while the subsequent evolution of the infra-orders and families has entailed a high level of chromosome variability only in colubroids. The karyological evolution in reptiles generally entailed a progressive reduction in chromosome changing rate, albeit with differences in the diverse orders and suborders. This trend seems to be consistent with the "canalization model" as originally proposed by Bickham and Baker in [Bickham, J.W. & R J. Baker, 1979. Bull. Carnegie Mus. Nat. Hist. 13: 70-84.] However, several inconsistencies have been found excluding that in this class the ultimate goal of chromosome variations was the achievement of a so-called "optimum karyotype'' as suggested by the above-mentioned theory. Other mechanisms could underpin chromosome variability in Reptiles. Among them a genomic composition more or less favourable to promoting chromosome rearrangements and factors favouring the fixation of a mutant karyotype in condition of homozygosis. Turtles and crocodiles would have a genome characterised by large chromosomes and a low level of chromosome compartmentalisation limiting the recombination and the frequency of rearrangements. A low rate of chromosome variability modifying little if at all the gene linkage groups would have favoured a conservative evolutionary strategy. In the course of evolution, lizards and snakes could have achieved a genome characterised by smaller chromosomes and a higher level of compartmentalisation. This would have raised the frequency of recombination and consequently an evolutionary strategy promoting a higher degree of variability and a greater level of speciation.

Animals↗
PubMed1998

Postmortem tracing reveals the organization of hypothalamic projections of the suprachiasmatic nucleus in the human brain.

The suprachiasmatic nucleus (SCN) is a small structure considered to be the site of the major circadian pacemaker of the mammalian brain. Disturbances in human biological clock function may occur in several diseases, such as Alzheimer's disease, sleep problems, and seasonal depression. Since basic knowledge of the anatomical connections of the human SCN is limited due to the lack of suitable neuroanatomical tracing methods, the understanding of physiological mechanisms of human SCN function has obviously been hampered. In the present study, the hypothalamic connections of the human SCN were revealed for the first time with a newly developed in vitro postmortem anterograde tracing method. The human SCN was found to be connected with nuclei in the hypothalamus that are involved in hormone secretion, cardiovascular regulation, and behavior activity. These human SCN projections appear to follow the same general patterns as those in the rodent brain. This homology may indicate an evolutionary conservation of the SCN projections from rodent to human. Through these connections, the human SCN may transmit its circadian information to regulate hormone secretion, body temperature, and behavioral functions as it does in animal species. In addition, the postmortem tracing technique may be a valuable tool that will contribute to our understanding of anatomical connections in the human brain, and may have other applications in the research on the physiology and pathology of the human brain.

Animals↗
PubMed1996

Yeast protein translocation complex: isolation of two genes SEB1 and SEB2 encoding proteins homologous to the Sec61 beta subunit.

A yeast gene (cDNA clone) was isolated in a screen for suppressors of secretion-defective sec15-1 mutation. This gene encodes a protein homologous to the beta subunit of the mammalian Sec61 protein complex functioning in protein translocation into the endoplasmic reticulum (ER). The predicted protein, Seb1p, consists of 82 amino acids and contains one potential membrane-spanning region at the C-terminus but no N-terminal signal sequence. Seb1p shows 30% identity to the mammalian Sec61 beta subunit and 34% identity to the Arabidopsis thaliana Sec61 beta subunit. Overexpression of SEB1 from a multicopy plasmid suppressed the temperature sensitivity of sec61-2 and sec61-3 mutants. Immunofluorescence and immunoelectron microscopy indicated that Seb1p resides in the ER membranes with the hydrophilic N-terminus exposed to the cytoplasm. The in vitro translated Seb1p was post-translationally inserted into microsomal membranes. As the chromosomal disruption of the SEB1 gene was not lethal, potential homologous genes were screened by heterologous hybridization. The SEB1 homologue thus isolated, SEB2, encodes a protein 53% identical to Seb1p. Disruption of the chromosomal SEB2 was not lethal whereas the double disruption of SEB1 and SEB2 resulted in a temperature-sensitive phenotype. This study further emphasizes the evolutionary conservation of the ER protein translocation apparatus and provides novel genetic tools for its functional analysis.

Amino Acid Sequence↗
PubMed1996

Cloning and expression of Xenopus CCT gamma, a chaperonin subunit developmentally regulated in neural-derived and myogenic lineages.

The chaperonin containing TCP-1 (CCT) is a eukaryotic cytoplasmic chaperonin, consisting of multiple distinct subunits in a double-toroid structure. In vitro, the CCT has been shown to assist in the folding of tubulin and actin into active conformations through an ATP-dependent mechanism. The function and distribution of these proteins in vivo are also not known. In this report, we show that the expression of two CCT subunits (alpha and gamma) are developmentally regulated in neural-derived and myogenic lineages. While expression in the central nervous system and muscle is consistent with a role in tubulin and actin conformation, we also detect robust expression in the developing cranial neural crest. Enrichment in the neural crest may represent the presence of a novel substrate for the CCT. We have also cloned the complete cDNA for the Xenopus ortholog of CCT gamma, which has 87% amino acid identity with the mouse protein. This remarkable evolutionary conservation suggests a conserved function for this protein among vertebrates, and possibly among all eukaryotes.

Amino Acid Sequence↗
PubMed1999

Functional analyses of a unique p53 germline mutant (Y236delta) associated with a familial brain tumor syndrome.

We have evaluated the functional properties of the unique p53 mutant Y236delta (deletion of codon 236) that gave rise to apparent cell-type specific tumor development. Four family members carrying this mutation in the germline developed early onset brain tumors, as previously reported. Deletion of residue Y236, which is tightly packed in an evolutionary conserved hydrophobic pocket, results in a protein with a mutant conformation according to immunoprecipitation with the conformation-sensitive antibodies PAb240 and PAb1620. The Y236delta mutant lacks specific DNA binding to the p53-responsive element in the WAF1-promoter, and functional analysis in Saos-2 cells revealed inability to transactivate the p53-responsive elements in the WAF1-promoter and the RGC sequence. The mutant has retained a functional oligomerization domain, a key element mediating the dominant negative effect, and inhibits DNA binding of wild-type p53. In addition, transactivation of endogenous wild-type p53 in LoVo cells was inhibited upon transfection of the mutant in a dose-dependent manner. Thus, in vitro and in vivo data suggest the loss of important tumor-suppressing functions and demonstrate a dominant negative effect of this unique p53 mutant that is associated with an unusual clustering of familial brain tumors.

Brain Neoplasms↗
PubMed1998

Novel miniproteins engineered by the transfer of active sites to small natural scaffolds.

Small multidisulfide-containing proteins are attractive structural templates to produce a biologically active conformation that mimics the binding surface of natural large proteins. In particular, the structural motif that is evolutionary conserved in all scorpion toxins has a small size (30-40 amino acid residues), a great structural stability, and high permissiveness for sequence mutation. This motif is composed of a beta-sheet and an alpha-helix bridged in the interior core by three disulfides. We have used this motif successfully to transfer within its beta-sheet new functional sites, including the curaremimetic loop of a snake neurotoxin and the CDR2-like site of human CD4. Accumulated evidence indicated that the two miniproteins produced, the curaremimetic miniprotein and the CD4 mimetic, contain the alpha/beta fold that is characteristic of the scaffold used and bind respectively to the acetylcholine receptor and to the envelope gp120 of HIV-1. Furthermore, the latter was shown to prevent viral infection of lymphocytes. These examples illustrate that, by the transfer of active sites to small and stable natural scaffolds, it is possible to engineer miniproteins reproducing, in part, the function of much larger proteins. Such miniproteins may be of great utility as tools in structure-function studies and as leads in drug design.

Amino Acid Sequence↗
PubMed1996

Tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA is constitutively expressed in bovine, human normal, and osteoarthritic articular chondrocytes.

Tissue inhibitors of metalloproteinases (TIMPs) inhibit the extracellular matrix (ECM) metalloproteinases (MMPs). To determine the source of TIMPs in synovial fluids of patients with osteoarthritis (OA), the ability of chondrocytes to express TIMP-2 and its regulation by agents found in inflammed joints was investigated. The constitutive TIMP-2 mRNA expression was demonstrated in chondrocytes from normal bovine, human OA and normal cartilage. The cross-hybridization of human and bovine TIMP-2 suggested its evolutionary conservation. Serum, IL-1, IL-6 and TGC-beta were unable to augment considerably the basal expression of TIMP-2 mRNA. TIMP-1 RNA expression in chondrocytes from human OA cartilage was elevated compared to non-OA chondrocytes, while TIMP-2 mRNA levels were similar in both. IL-1 beta, IL-6 and TGF-beta did not affect TIMP-2 expression but TGF-beta induced TIMP-1 mRNA in human OA chondrocytes. TIMP-2 and TIMP-1 are therefore differentially regulated in chondrocytes and the basal TIMP-2 levels may be needed for the cartilage ECM integrity.

Animals↗
PubMed1998

Molecular cloning and tissue-specific expression of Mrad9, a murine orthologue of the Schizosaccharomyces pombe rad9+ checkpoint control gene.

We have isolated a murine cDNA, Mrad9, that is orthologous to the fission yeast rad9+ and human HRAD9 genes. Mrad9 encodes a 389 amino acid long, 42,032 Dalton protein that is 27% identical and 56% similar to Rad9p, and 82% identical and 88% similar to HRAD9, at the amino acid level. Expression of the Mrad9 cDNA in Schizosaccharomyces pombe rad9::ura4+ cells restores nearly wild-type levels of hydroxyurea resistance and early S phase checkpoint control to mutant fission yeast cell populations. However, UV resistance is only minimally restored, and mutant cells remain sensitive to gamma radiation. Mrad9 genomic DNA was isolated from a mouse 129/SvEv library. The Mrad9 gene was local ized to a 15-kbp genomic DNA fragment, and contains 10 exons separated by 9 introns. Northern blot analysis indicates that the gene is expressed in many different tissues of the adult mouse, but the mRNA is most abundant in the heart and present at very low levels in the liver. These studies demonstrate the existence of a murine orthologue of the fission yeast rad9+ gene and underscore at least the partial evolutionary conservation of rad9+-dependent checkpoint control mechanisms.

Amino Acid Sequence↗
PubMed1997

Identification of 16 sulfamidase gene mutations including the common R74C in patients with mucopolysaccharidosis type IIIA (Sanfilippo A).

Mucopolysaccharidosis type IIIA (MPS IIIA or Sanfilippo A disease) is a storage disorder caused by deficiency of the lysosomal enzyme sulfamidase. Mutation screening, using SSCP/heteroduplex analyses on cDNA and genomic DNA fragments, was performed in a group of 42 European patients. Sixteen of the 17 different gene mutations characterized have not been previously described. The spectrum of gene lesions consists of two 1-bp deletions (1091delC, 1093delG), an 18-bp duplication (421ins18), a splice site mutation (IVS2-2A-->G), and 13 different missense point mutations. As in other lysosomal storage disorders, the phenotypic heterogeneity is associated with a considerable genetic heterogeneity. The missense mutation R74C, which alters an evolutionary conserved amino acid in the active site of the enzyme, was found on 56% of alleles of 16 Polish patients, whereas it was less frequent among German patients (21% of disease alleles). R245H, a previously reported common mutation, represents 35% of disease alleles in German patients, but only 3% in Polish patients. As the combined frequency of the common mutations (R74C and R245H) in German and Polish populations exceeds 55%, screening for these two mutations will assist molecular genetic diagnosis of MPS IIIA and allow heterozygote testing in these populations.

Binding Sites↗
PubMed1998

Further delineation of the molecular pathology of Wilson disease in the Mediterranean population.

This study presents the update results of an ongoing project on the delineation of the spectrum of mutations at the Wilson disease (WD) gene in WD patients of Mediterranean origin. In studying 59 patients, of whom were 26 Continental Italians, 22 Sardinians, 9 Turkish, and 2 Albanians, we have found 31 novel and three known mutations. Of the novel mutations, 3 are deletions, two nonsense, 2 splice or consensus splice site, and 24 missense. The large majority of the missense mutations lie in evolutionary conserved regions of the WD gene of documented functional importance. Most of our patients were compound heterozygotes, and only a few were homozygotes. In addition, three polymorphisms were detected. By adding the new data to those previously reported by our group, we have to date detected 85% of mutations in the WD chromosomes from Continental Italians, 30% from Sardinians, 81.7% from Turkish and 66.7% from Albanians. Most of the mutations characterized are rare, and only a limited number are common. Of the common mutations 5 were found in Continental Italians, two in Sardinians and a single one in Turkish. Because there are so many causative mutations of the disease, the preclinical and prenatal diagnosis of WD should be carried out by a combination of mutation and linkage analysis.

Adenosine Triphosphatases↗
PubMed1998

Combinatorial interactions regulating cardiac transcription.

In vertebrates, heart development is a multistep process that starts with formation and patterning of the primitive heart tube and is followed by complex morphological events to give rise to the mature four-chambered heart. These various stages are characterized by distinct patterns of gene expression. Although chamber specificity and developmental regulation can be demonstrated in transgenic mice using short promoter fragments, the mechanism underlying spatial and temporal specificity within the heart remains largely unclear. Combinatorial interaction between a limited number of cardiac-specific and ubiquitous transcription factors may account for the diverse genetic inputs required to generate the complex transcriptional patterns that characterize the developing myocardium. We have used the cardiac atrial natriuretic peptide (ANP) promoter to test this hypothesis. The ANP gene is transcribed in a spatial- and temporal-specific manner in the heart, and a 500 bp promoter fragment is sufficient to recapitulate both chamber and developmental specificity. This promoter is composed of three modules, a "basal" cardiac promoter that is essential for transcription in embryonic and postnatal atrial and ventricular myocytes and two other independent modules that behave as chamber-specific enhancers. The basal cardiac promoter is the target of two cardiac-specific transcription factors, the zinc finger GATA-4 protein and the Nkx2-5 homeodomain, which bind to contiguous elements within this region. At low concentrations--a situation that likely occurs during the very first stages of cardiac cell fate determination--the two proteins synergistically activate transcription from the ANP promoter. This functional synergy requires physical interaction between the GATA-4 protein and an extended C-terminal homeodomain on Nkx2-5. This interaction, which unmasks an activation domain present just N-terminal of the homeodomain, is specific for GATA-4 and-5, but is not observed with the other cardiac GATA factor, GATA-6. Optimal synergy requires binding of both proteins to their cognate sites, although modest synergy also could be observed on heterologous promoters containing only multimerized Nkx binding sites, suggesting that Nkx2-5 is able to recruit GATA-4 into a transcriptionally active complex. The GATA/Nkx interaction, which appears to have been evolutionary conserved in nematode, fly, and mammals, provides a paradigm for analyzing transcription factor interaction during organogenesis. The data are also discussed in the context of our present knowledge of the roles of GATA and NK2 proteins in cardiac development.

Animals↗
PubMed2000

Differential expression of the PSD-95 gene family in electrosensory neurons.

The PSD-95 family of membrane-associated guanylate kinase (MAGUK) proteins are involved in the assembly and organization of neurotransmitter receptors at excitatory synapses in the vertebrate nervous system. We have isolated partial cDNAs for five PSD-95 family members from Apteronotus leptorhynchus brain RNA using a degenerate PCR method. The amino acid sequences deduced indicate that A. leptorhynchus neurons express homologues of the mammalian PSD-93, SAP-97, and SAP-102 MAGUKs and two homologues of mammalian PSD-95. In situ hybridization experiments have been carried out to localize the cellular expression of all five MAGUK mRNAs in the central nervous system of A. leptorhynchus. In the cerebellum the expression patterns are highly similar to patterns reported for mammalian cerebellum, suggesting an evolutionary conservation of the functional roles in this gene family. Cellular levels of expression of the PSD-95 MAGUK mRNAs and the NMDAR-1 mRNA were highly correlated in neurons of the dorsal forebrain but were not correlated in neurons of the electrosensory lateral line lobe (ELL) or the cerebellum. These results suggest that the expression of PSD-95 MAGUK genes in forebrain neurons may provide mechanisms for synaptic organization that are not shared by neurons in the ELL and cerebellum.

Amino Acid Sequence↗
PubMed1993

Mutations of p53 and human papillomavirus infection in cervical carcinoma.

BACKGROUND: Oncogenic human papillomavirus (HPV) infection has been implicated in the pathogenesis of cervical carcinoma. The HPV oncoproteins E6 and E7 are though to play a crucial role in this process by their interactions with the p53 protein and the retinoblastoma susceptibility gene product, respectively. The E6 protein binds to and stimulates the degradation of the p53 protein. Mutations involving evolutionary conserved regions of the p53 gene also can alter p53 function. Point mutations of p53 frequently have been identified in a wide variety of human tumors. METHODS: Forty-five cervical carcinoma samples were evaluated for the presence of mutations involving exons 5-8 of the p53 gene with polymerase chain reaction (PCR) amplification of genomic DNA, followed by single-stranded conformation polymorphism analysis and/or direct sequencing. The status of oncogenic HPV infection in the tumor tissues was analyzed by Southern blot and PCR. RESULTS: Forty-two of 45 cervical carcinomas showed oncogenic HPV DNA: Of three HPV-negative samples, one harbored a missense point mutation of the p53 gene. An additional p53 point mutations was identified in a tumor with HPV 18 infection. CONCLUSIONS: Oncogenic HPV DNA can be identified in most cervical carcinomas. Mutations involving conserved regions of p53, although infrequent in cervical cancer, occur preferentially in tumors without HPV infection. Inactivation of p53 function is important in the pathogenesis of cervical carcinoma.

Adenocarcinoma↗
PubMed2000

Identification of four single nucleotide polymorphisms in DNA repair genes: XPA and XPB (ERCC3) in Polish population.

A deficiency in DNA repair is associated with increased cancer risk. Inter-individual variations in DNA repair capacity observed in humans may result from genetic polymorphisms in DNA repair genes. In order to provide a basis for future functional and molecular epidemiology studies on cancer susceptibility, we screened 35 individuals for polymorphisms in coding regions of XPA and XPB genes involved in nucleotide excision repair (NER). Relevant cDNA sequences were amplified by PCR, sequenced with fluorescently labeled terminators and analyzed with automated sequencer. Two polymorphisms in XPB were found: AAA-->AGA (445A>G; GenBank M31899) causing K117R substitution and GGC-->TGC (1299G>T; GenBank M31899) causing G402C exchange. Also, two polymorphisms in XPB were detected: CGA-->CAA (709G>A; GenBank D14533) causing R228Q exchange, and A-->G (23A>G; GenBank D14533) substitution in the 5' non-coding region of the gene. The three aforementioned amino acid substitutions were uncommon in this population (1.4%). In contrast, the substitution located 4 nucleotides upstream of the ATG start codon of XPB was frequent (57%). To our best knowledge this is the first report of these sequence variants. The location of these polymorphisms in evolutionary conserved regions suggest that they may be of functional significance.

Animals↗
PubMed1987

Epigenetic rules for expression of cell adhesion molecules during morphogenesis.

From very early developmental times, cell adhesion molecules (CAMs) play key roles in linking cells together and regulating cell movement. By virtue of their capacity to link epithelia and condense mesenchyme, CAMs can act as mechanochemical regulators of morphogenesis. In the vertebrate species examined so far, CAMs appear in ordered sequences on cell surfaces during development. In this paper, evidence is reviewed indicating that the sequential expression of CAMs on cell surfaces at a variety of sites of embryonic induction follows a set of modulation rules that are first discernible at early gastrulation. These rules are related to the adhesion of cells in collectives and to the establishment of borders between such collectives. After gastrulation, all mesenchymal conversions employ N-CAM and show changes in its prevalence in a transition N----0----N where 0 means low or undetectable amounts of the CAM (rule I). In contrast, epithelia modulate from a state in which N-CAM and L-CAM appear simultaneously to the expression of only one or the other of these primary CAMs (rule II). At a variety of induction sites, cell collectives obeying rule I are found in proximity to cell collectives obeying rule II. During the morphogenesis of complex structures such as the feather or the optic placode, one can see a recursive application of these rules, reflecting the formation of significant histological boundaries within which the expression of gene products other than CAMs can lead to great morphological diversity. It is suggested that the genes for CAMs are regulated independently from and prior to those specifying intracellular proteins in a given tissue. According to this proposal, the existence of the epigenetic rules governing CAM expression reflects the evolutionary conservation of a key means of establishing tissue and animal form through the mechanochemical regulation of processes such as cell division, movement and death.

Animals↗
PubMed1989

The cellular interphase.

A brief outline is given of the impact of molecular genetics on the study of morphogenesis. Two approaches have been used, one beginning at the molecular level and applying knowledge gained in this way to observations of whole organisms; the other starting with descriptions of the organism and tracing the causes of developmental changes to cellular and then molecular events. The availability of experimentally induced mutants has facilitated the study of development, but such work must be combined with detailed information on the normal process. Comparative studies have exploited evolutionary conservation to identify functionally important gene activities, and allowed knowledge gained from one organism to be rapidly applied to other species. The successes achieved at the molecular level are impressive, but the time has come to return to studying the cells that are the real mediators of the transmission of the genetic information.

Animals↗
PubMed1983

Hormonal regulation and expression of vitellogenin multigene family.

Yolk proteins are the most abundant egg proteins in oviparous animals. They are deposited during oocyte maturation for use after fertilization and are synthesized in the liver or fat body as a common precursor termed vitellogenin. Hybridization with cloned DNA complementary to vitellogenin messenger RNA has revealed a surprisingly high degree of evolutionary conservation of sequence of vitellogenin genes among insects, amphibians and birds. The synthesis of vitellogenin in vertebrates is directly under the control of oestrogen at the level of gene transcription. In the frog, Xenopus, vitellogenin genes occur as a multigene family, four of which are actively expressed and are grouped as A and B genes. This multiplicity offers a useful system for investigating the possible selective hormonal regulation of expression of individual members of multigene families. When X. laevis vitellogenin genes were activated by oestrogen in the liver of whole animals or in cultures of parenchymal cells, the two groups of expressed genes were not induced in an identical manner in cells from male and female animals. The activation of A and B groups of genes was non-coordinate in male hepatocytes and coordinate in female cells. Prior exposure of male hepatocytes to oestradiol in vivo or in culture caused the pattern of expression to shift to that in female cells. Since the X. laevis oocyte itself does not synthesize vitellogenin in response to oestrogen, an attempt was made to activate its dormant vitellogenin genes by transferring oestrogen-binding proteins from the liver. Preliminary results show that the microinjection into the oocyte of a preparation containing liver receptor-hormone complex led to the synthesis of vitellogenin by the oocyte. Extension of these experiments will not only enable a more precise analysis of the activation of the vitellogenin multigene family to be made but will also provide direct functional evidence for the role played by steroid hormone receptors in regulating gene expression.

Animals↗
PubMed1985

Receptors for the phorbol ester tumour promoters.

The phorbol esters are potent tumour promotors in mouse skin and have profound effects on a wide variety of biological systems. Using the derivative [20-3H]phorbol 12,13-dibutyrate ([3H]PDBu), we demonstrated that specific receptors for the phorbol esters exist which mediate many of these biological effects. The receptors represent a complex between phospholipids and protein kinase C, an enzyme first identified by Nishizuka and coworkers. Considerable evidence indicates heterogeneity in the pharmacology of the biological responses to the phorbol esters. Likewise, multiple subclasses of binding sites have been observed. Differences in the phospholipids associated with protein kinase C may account for this heterogeneity, affecting both absolute and relative affinities. An endogenous analogue of the phorbol esters had been predicted from the high evolutionary conservation of the receptor. Nishizuka and coworkers have reported that diacylglycerols appear to be natural activators of protein kinase C. We find that diacylglyerols competitively inhibit phorbol ester binding, consistent with their acting at the same site on the enzyme. Likewise, by binding analysis, we can demonstrate a 1:1 stoichiometry between D-1-2-diacylglycerol and the receptor. The potencies of diacylglycerols for binding reflect their local concentration in the phospholipids. The K1 for diolein is approximately 0.1%. Relative to the corresponding phorbol esters, the potencies of the diacylglycerols are somewhat (17-fold) to markedly (30 000-fold) lower. We conclude that factors affecting phospholipid-receptor interactions or diacylglycerol production are of potential importance in the promotion process.

Animals↗