21 citations found

Insect Biochem Mol Biol 27 (1): 27-35 (1997)

Complete nucleotide sequence of the vitellogenin mRNA from the gypsy moth: novel arrangement of the subunit encoding regions.

Hiremath S, Lehtoma K

USDA Forest Service Northeastern Forest Experiment Station, Delaware, OH 43015, USA.

Primary structure analysis and location of introns suggests evolutionary relatedness among vitellogenin (Vg) genes from vertebrates and invertebrates, including insects. We have determined the complete nucleotide sequence of the gypsy moth VgmRNA, which shows that its structure is significantly different from VgmRNAs in other systems. The nucleotide sequence was determined using overlapping cDNA fragments generated from RACE reactions and rTh polymerase-mediated PCR. The VgmRNA is 5579 nucleotides long and codes for both the large and small subunits. However, the arrangement of the subunit encoding regions in the gypsy moth VgmRNA is opposite of what has been observed in other systems. Gypsy moth Vg gene is the first reported example of a Vg gene where the 5'-terminal region codes for the large subunit and the 3'-terminal region for the small subunit. Also, the sequence near the junction of subunits was significantly different from those found in other insects. This may be responsible for the relatively more stable precursor of Vg subunits found in the gypsy moth hemolymph. It is not clear where this divergence in the structure of Vg gene occurred during evolution, since the Vg gene of another lepidopteran, Bombyx mori, conforms to the structure of those in vertebrates and other invertebrates. 


J Mol Evol 41 (4): 505-521 (1995)

Fundulus heteroclitus vitellogenin: the deduced primary structure of a piscine precursor to noncrystalline, liquid-phase yolk protein.

LaFleur GJ Jr, Byrne BM, Kanungo J, Nelson LD, Greenberg RM, Wallace RA

Whitney Laboratory, University of Florida, Marineland 32086, USA.

We have cloned and sequenced a cDNA encoding a vitellogenin (Vtg) from the mummichog, Fundulus heteroclitus, an estuarine teleost. We constructed a liver cDNA library against RNA from estrogen-treated male mummichogs. Five overlapping cDNA clones totalling 5,197 bp were isolated through a combination of degenerate oligonucleotide probing of the library and PCR. The cDNA sequence contains a 5,112 bp open reading frame. The predicted primary structure of the deduced 1,704-amino-acid protein is 30-40% identical to other documented chordate Vtgs, establishing this Vtg as a member of the ancient Vtg gene family. Of the previously reported chordate Vtg sequences (Xenopus laevis, Gallus domesticus, Ichthyomyzon unicuspis, and Acipenser transmontanus), all four act as precursor proteins to a yolk which is eventually rendered insoluble under physiological conditions, either as crystalline platelets or as noncrystalline granules. The yolk of F. heteroclitus, on the other hand, remains in a soluble state throughout oocyte growth. The putative F. heteroclitus Vtg contains a polyserine region with a relative serine composition that is 10-20% higher than that observed for the other Vtgs. The trinucleotide repeats encoding the characteristic polyserine tracts of the phosvitin region follow a previously reported trend: TCX codons on the 5' end and AGY codons toward the 3' end. Whether the difference in Vtg primary structure between F. heteroclitus and that of other chordates is responsible for the differences in yolk structure remains to be elucidated. As the first complete teleost Vtg to be reported, these data will aid in designing nucleotide and immunological probes for detecting Vtg as a reproductive status indicator in F. heteroclitus and other piscine species. 


J Mol Evol 34 (6): 478-492 (1992)

The boll weevil vitellogenin gene: nucleotide sequence, structure, and evolutionary relationship to nematode and vertebrate vitellogenin genes.

Trewitt PM, Heilmann LJ, Degrugillier SS, Kumaran AK

Biology Department, Marquette University, Milwaukee, WI 53233.

Boll weevil (Anthonomus grandis) eggs contain two yolk proteins, YP47 and YP160. Using anti-YP160 antiserum as probe, a partial-length complementary DNA (cDNA) was isolated from a lambda gt11 adult female cDNA library. A second partial-length cDNA was isolated from a lambda gt10 adult female cDNA library by differential screening with male vs. female cDNAs. Northern blot analysis showed that each cloned cDNA hybridized to a 6-kb female-specific transcript. These cDNAs were used to probe a genomic library, and two overlapping genomic clones were obtained that span the boll weevil vitellogenin gene. The entire transcription unit was sequenced, and introns were mapped by a combination of primer extension experiments, S1 nuclease protection experiments, and polymerase chain reaction-mediated synthesis of two additional cDNA clones. Based on these data, the vitellogenin mRNA is 5511 nucleotides [plus a poly(A) tail of undetermined length] and specifies a provitellogenin of 1790 amino acids. The deduced protein has a Glu+Gln content of 16.3%, which is a relatively high value that is typical of most vitellogenins. Protein sequence similarities including Cys clusters conserved between boll weevil vitellogenin and Xenopus laevis A2 or Caenorhabditis elegans vit-5 vitellogenins indicated that the boll weevil protein is a member of the ancient nematode-vertebrate vitellogenin family. Moreover, the six introns in the boll weevil vitellogenin gene interrupt the coding region at positions closely or exactly corresponding to a subset of the positions of the 34 vertebrate vitellogenin introns, further supporting the argument for a common evolutionary relationship. This report represents the first complete nucleotide sequence and structural analysis of a nondipteran insect vitellogenin gene. 


J Lipid Res 33 (6): 777-790 (1992)

Why is there sequence similarity between insect yolk proteins and vertebrate lipases?

Bownes M

Institute of Cell and Molecular Biology, University of Edinburgh.

The major proteins stored in the yolk of developing oocytes are thought to provide a nutritional store for utilization during embryogenesis. They seem to fall into two major families of proteins. The first are called vitellogenins and are found in frog, chicken, nematode, fish, and some insects such as the boll weevil. The other group are called yolk proteins and are found in dipteran insects such as fruitfly, housefly, fleshfly, and blue-bottles. Both groups are the major proteins found in the oocyte and are female-specific proteins endocytosed from the serum or hemolymph. The yolk protein group were found to have sequence similarity to the triacylglycerol lipases and lipoprotein lipases of vertebrates, including rat, pig, and human. The yolk proteins do not have lipase activity, but the sequences conserved between yolk proteins and lipases surround the active site where there are interactions with lipids. The likely reason for the presence of this domain in the yolk proteins is to bind a steroid hormone in a storage form conjugated to lipids. This permits the storage of the hormone in an inactive form until the yolk proteins are degraded, when it can be released from its conjugate to induce developmental decisions in embryogenesis. They may also transport lipids into the oocyte for use in embryogenesis. Whilst the vitellogenin family of proteins do not share this homology with the lipases they do have similarity to the human serum protein, apolipoprotein B, which also has a role in binding lipids. These findings are discussed in relation to the evolution and functions of lipases, apolipoproteins, vitellogenins, and yolk proteins. Experiments aimed at isolating genes encoding lipases in insects and at further elucidating the function of the yolk proteins are suggested. 


Genetics 127 (4): 769-780 (1991)

A cluster of vitellogenin genes in the Mediterranean fruit fly Ceratitis capitata: sequence and structural conservation in dipteran yolk proteins and their genes.

Rina M, Savakis C

Institute of Molecular Biology and Biotechnology, Foundation of Research and Technology, Heraklion, Crete, Greece.

Four genes encoding the major egg yolk polypeptides of the Mediterranean fruit fly Ceratitis capitata, vitellogenins 1 and 2 (VG1 and VG2), were cloned, characterized and partially sequenced. The genes are located on the same region of chromosome 5 and are organized in pairs, each encoding the two polypeptides on opposite DNA strands. Restriction and nucleotide sequence analysis indicate that the gene pairs have arisen from an ancestral pair by a relatively recent duplication event. The transcribed part is very similar to that of the Drosophila melanogaster yolk protein genes Yp1, Yp2 and Yp3. The Vg1 genes have two introns at the same positions as those in D. melanogaster Yp3; the Vg2 genes have only one of the introns, as do D. melanogaster Yp1 and Yp2. Comparison of the five polypeptide sequences shows extensive homology, with 27% of the residues being invariable. The sequence similarity of the processed proteins extends in two regions separated by a nonconserved region of varying size. Secondary structure predictions suggest a highly conserved secondary structure pattern in the two regions, which probably correspond to structural and functional domains. The carboxy-end domain of the C. capitata proteins shows the same sequence similarities with triacyglycerol lipases that have been reported previously for the D. melanogaster yolk proteins. Analysis of codon usage shows significant differences between D. melanogaster and C. capitata vitellogenins with the latter exhibiting a less biased representation of synonymous codons. 


Biochem Genet 28 (7-8): 415-432 (1990)

Vitellogenin protein diversity in the Hawaiian Drosophila.

Craddock EM, Kambysellis MP

Division of Natural Sciences, State University of New York, Purchase 10577.

Egg and female hemolymph proteins were resolved via SDS-polyacrylamide gel electrophoresis in a diverse array of 33 endemic Hawaiian drosophilids, encompassing 17 picture-winged species, 3 of the antopocerus species group, 9 fungus feeders, 1 species from each of the modified mouthparts, crassifemur and ciliated tarsus groups, and 1 Scaptomyza species. Molecular weights of the two (10 species) or three vitellogenin bands (22 species) were highly variable, spanning a 7-kD range. The largest vitellogenin, V1, was the most variable, showing a change of some 10% in its mean size of 47.6 kD. The smallest V3 vitellogenin, mean size 44.1 kD, was evolutionarily the most conservative in size. The species Drosophila hawaiiensis was found to be polymorphic for two/three vitellogenin bands and, also, polymorphic with respect to the size of the V1 protein. No inter- or intrapopulation variability in vitellogenin size was detected in 10 other species examined. The major features of vitellogenin protein evolution in the Hawaiian Drosophila are change in molecular weight and regulatory differences that result in quantitative differences between species in patterns of vitellogenin protein production. 


J Mol Evol 28 (6): 487-496 (1989)

Potential regulatory elements of nematode vitellogenin genes revealed by interspecies sequence comparison.

Zucker-Aprison E, Blumenthal T

Department of Biology, Indiana University, Bloomington 47405.

The nematode, Caenorhabditis elegans, has a six-member gene family encoding vitellogenins, the yolk protein precursors. These genes are expressed exclusively in the intestine of the adult hermaphrodite. Here we report the cloning of all five members of the homologous gene family from another Caenorhabditis species, Caenorhabditis briggsae. Nucleotide sequence analysis of these genes reveals they are about 85% identical to the C. elegans genes in the coding regions. Overall similarity is much reduced in noncoding and flanking regions. However, two repeated heptamers, previously identified in the upstream regions of the C. elegans genes, are largely conserved in both location and sequence in C. briggsae. Conservation of certain of these heptamers suggests that proteins bound at these positions may be especially important to promoter function and/or regulation. Comparative sequence analysis also suggests the possibility that the first 70 bases of the vitellogenin mRNAs can be folded into stable secondary structures. Almost all base differences between the two species occur in sequences predicted to be unpaired, suggesting that the ability to form intrastrand base pairs has been selected during Caenorhabditis evolution. 


Prog Biophys Mol Biol 53 (1): 33-69 (1989)

The evolution of egg yolk proteins.

Byrne BM, Gruber M, Ab G




Biochem J 255 (3): 1057-1060 (1988)

Is vitellogenin an ancestor of apolipoprotein B-100 of human low-density lipoprotein and human lipoprotein lipase?

Baker ME

Department of Medicine, University of California, San Diego, La Jolla 92093.

Vitellogenin, an ancient animal protein, is the major yolk protein of eggs, where it is used as a food source during embryogenesis. Here it is shown that vitellogenins, including those from the invertebrates Caenorhabditis elegans and Drosophila melanogaster, contain domains that are homologous with parts of apolipoprotein B-100 (apoB-100) of human low-density lipoprotein and human lipoprotein lipase. As vitellogenins are likely to have been used by invertebrates during embryogenesis well before the circulation of lipids appeared in vertebrates, it is suggested that copies of a precursor gene, serving a function similar to vitellogenin, were modified to code for part of apoB-100 and lipoprotein lipase in vertebrates. In addition to providing a link between invertebrates and vertebrates for proteins involved in lipid transport, these homologies suggest new functions for vitellogenin other than being a yolk food for the developing embryo. 


Trends Genet 4 (8): 227-232 (1988)

Evolution and expression of vitellogenin genes.

Wahli W




C R Acad Sci III 306 (14): 441-446 (1988)

Evolution of vitellins in the ovary and during development in three species of Diptera, Drosophila melanogaster, Calliphora vicina and Prasarcophaga argyrostoma

Karlinsky A, Mendes H, Schoeller-Raccaud J, Thomas-Orillard M

Laboratoire de Physiologie des Insectes, Universite Pierre-et-Marie-Curie, Paris.

The SDS electrophoresis reveals the asynchronous appearance of the three vitellins V1, V2, V3 during the ovarian cycle. It is always the V3 which appears first with the beginning of previtellogenesis. The vitellin degradation occurs late at the end of embryogenesis. The presence of a protein migrating at the level of V3 during the whole development is discussed. 


Nucleic Acids Res 15 (24): 10405-10417 (1987)

A single gene encoding vitellogenin in the sea urchin Strongylocentrotus purpuratus: sequence at the 5' end.

Shyu AB, Blumenthal T, Raff RA

Institute for Molecular and Cellular Biology, Indiana University, Bloomington 47405.

The synthesis of vitellogenin (yolk protein precursor) in the sea urchin, Strongylocentrotus purpuratus, is unique in that both males and females produce a high level of the protein. In this paper we show that this organism also is unique in possessing only a single vitellogenin gene. Like the genes that encode analogous proteins in vertebrates, the sea urchin gene is large, about 19 kb in length. The sequence surrounding the 5' end of the gene revealed several other similarities to vertebrate vitellogenin genes: the signal sequence is exceptionally short and has a sequence similar to those from frog and chick; there is a canonical TATA box at -32; and there is a sequence closely resembling the estrogen-responsive element at -207. 


J Biol Chem 262 (32): 15377-15385 (1987)

Comparison of the organization and fine structure of a chicken and a Xenopus laevis vitellogenin gene.

Nardelli D, van het Schip FD, Gerber-Huber S, Haefliger JA, Gruber M, Ab G, Wahli W

Institut de Biologie Animale, Universite de Lausanne, Switzerland.

The structural organization and the coding nucleotide sequence of the Xenopus laevis A2 and the chicken major vitellogenin genes have been compared. Both genes show the same exon-intron organization. However, the degree of homology between the nucleotide and derived amino acid sequences varies extensively along the genes. Several of the 35 exons are quite similar, and a unique cysteine motif in the lipovitellin II domain is conserved between the two genes. In contrast, one internal region is quite divergent. Part of this region encodes phosvitin, which appears to have evolved rapidly by both point mutations and duplications of serines or short other amino acid stretches. On the basis of these observations, we discuss the possible mechanism of evolution of phosvitin in vertebrates. 


Biochemistry 26 (20): 6397-6402 (1987)

Vertebrate and nematode genes coding for yolk proteins are derived from a common ancestor.

Nardelli D, Gerber-Huber S, van het Schip FD, Gruber M, Ab G, Wahli W

Institut de Biologie animale, Universite de Lausanne, Switzerland.

One of the most obvious characteristics of the egg cells of oviparous animals is their large size resulting to a major extent from the deposition of nutritional reserves, mainly constituted of yolk proteins. In general, these are derived from a precursor called vitellogenin, which undergoes posttranslational modifications during secretion and during transport into and storage within the oocytes. Comparative analysis of the structural organization of the vitellogenin gene and of its product in different species shows that the vitellogenin gene is very ancient and that in vertebrates the gene may have more resemblance to the earliest gene than in invertebrates. 


Cell 46 (7): 1053-1061 (1986)

An estrogen-responsive element derived from the 5' flanking region of the Xenopus vitellogenin A2 gene functions in transfected human cells.

Klein-Hitpass L, Schorpp M, Wagner U, Ryffel GU

In the human breast cancer cell line MCF-7, we observe estrogen induction of the stable transfected Xenopus vitellogenin A2 gene. An estrogen-responsive element (ERE) could be defined by using a vitellogenin-chloramphenicol acetyltransferase hybrid gene in transient transfection experiments. The ERE is located in the 5' flanking region and is able to confer estrogen inducibility to the thymidine kinase gene promoter. By 5' and 3' deletions we have determined a 35 bp sequence sufficient for high stimulation by estradiol. Even 18 bp give a small estrogen response. The 35 bp ERE contains the palindromic sequence 5'GGTCACAGTGACC-3' as an essential element. The fact that the ERE of a frog gene functions in human cells demonstrates that signals and factors involved in the control have been conserved during evolution. 


Mol Cell Biol 5 (10): 2495-2501 (1985)

The Caenorhabditis elegans vitellogenin gene family includes a gene encoding a distantly related protein.

Spieth J, Blumenthal T

While the nematode Caenorhabditis elegans is more primitive than most egg-laying organisms, it's vitellogenins, or yolk protein precursors, appear to be more complex. C. elegans oocytes accumulate two major classes of yolk proteins. The first consists of two polypeptides with an Mr of about 170,000 (yp170A and yp170B) encoded by a family of five closely related genes called vit-1 through vit-5. The second class consists of two smaller proteins with Mr values of 115,000 (yp115) and 88,000 (yp88) which are cut from a single precursor. Here we report the cloning and analysis of a single-copy gene (vit-6) that encodes this precursor. The lengths of the gene and its mRNA are about 5 X 10(3) base pairs. Like vit-1 through vit-5, vit-6 is expressed exclusively in adult hermaphrodites. Comparison of portions of the coding sequence indicates that vit-6 is distantly related to the vit-1 through vit-5 gene family. Thus, even though the two classes of yolk proteins are antigenically and physically distinct, they are encoded by a single highly diverged gene family. 


Comp Biochem Physiol [B] 82 (3): 497-505 (1985)

Vitellogenin genes and their products in closely and distantly related species of Xenopus.

Baker BS, Steven J, Tata JR

Plasma vitellogenins from two closely related species of Xenopus, X. laevis and X. borealis, and a more ancient species, X. tropicalis, exhibited the same size on gel electrophoresis and were immunologically related. Partial peptide maps of 125I-labelled plasma vitellogenins, however, revealed marked differences in th structure and organisation of vitellogenin in the three Xenopus species. Northern blot hybridisation of liver RNA from oestrogen-treated males and females, probed with cloned vitellogenin cDNA, revealed the presence of mRNA of the same size in the three species of Xenopus, which was absent in untreated male liver. Cell-free translation of total liver RNA showed the presence of functional mRNA coding for vitellogenin subunit of the same size (Mr congruent to 210,000). Restriction endonuclease digestion patterns of genomic DNA from the three Xenopus species, using cloned X. laevis vitellogenin cDNA as the hybridisation probe, revealed significant differences in the organisation of these genes, which occur at a higher multiplicity in X. laevis and X. borealis than in X. tropicalis. Thus, despite a high degree of conservation of size, overall sequence and immunological identity of vitellogenin genes and their products in the three species of Xenopus, there is a substantial structural rearrangement during evolution of Xenopus within this multigene family. 


Nucleic Acids Res 12 (22): 8595-8609 (1984)

Evolution of vitellogenin genes: comparative analysis of the nucleotide sequences downstream of the transcription initiation site of four Xenopus laevis and one chicken gene.

Germond JE, Walker P, ten Heggeler B, Brown-Luedi M, de Bony E, Wahli W

Electron microscopic analysis of heteroduplexes between the most distantly related Xenopus vitellogenin genes (A genes X B genes) has revealed the distribution of homologous regions that have been preferentially conserved after the duplication events that gave rise to the multigene family in Xenopus laevis. DNA sequence analysis was limited to the region downstream of the transcription initiation site of the Xenopus genes A1, B1 and B2 and a comparison with the Xenopus A2 and the major chicken vitellogenin gene is presented. Within the coding regions of the first three exons, nucleotide substitutions resulting in amino acid changes accumulate at a rate similar to that observed in globin genes. This suggests that the duplication event which led to the formation of the A and B ancestral genes in Xenopus laevis occurred about 150 million years ago. Homologous exons of the A1-A2 and B1-B2 gene pairs, which formed about 30 million years ago, show a quite similar sequence divergence. In contrast, A1-A2 homologous introns seem to have evolved much faster than their B1-B2 counterparts. 


Ciba Found Symp 98: 96-110 (1983)

Hormonal regulation and expression of vitellogenin multigene family.

Tata JR, James TC, Watson CS, Williams JL, Wolffe AP

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. 


J. Mol Evol 18 (6): 405-413 (1982)

An evolutionary model for the insect vitellins.

Harnish DG, White BN

Insects can be divided into three groups based on the sizes of the polypeptide constituents of their vitellogenins and vitellins. In order to determine the relationships between these groups, antisera to the vitellins of seven insects from six taxonomic orders were used to assess immunological cross-reactivity. Antigenic relatedness was observed only between vitellins from species within the same family. Amino acid compositional data for vitellins from nine species were used to assess homology by difference matrices. The S delta Q values were similar for both intra-order and inter-order comparisons and strongly suggested relatedness. The S delta n comparisons supported the immunological data that indicated that the vitellins were evolving rapidly. For most insect vitellins there are two distinct size classes of polypeptides that seem to be derived from a single asymmetric proteolytic cleavage of a precursor. We propose a model that suggests that the different size polypeptides represent distinct domains and that in the evolution of the vitellogenin genes of the Diptera and Hymenoptera there has been domain elimination. 


DNA 1 (4): 345-353 (1982)

Evolutionary conservation of vitellogenin genes.

James TC, Bond UM, Maack CA, Applebaum SW, Tata JR

Homologous and heterologous hybridizations in solution were performed between sheared genomic DNA and DNA complementary to vitellogenin mRNA of Xenopus, chicken, and migratory locust. The kinetics of hybridization and the thermal stability of the hybrids formed suggested a high degree of conservation of coding sequences of insect, amphibian, and avian vitellogenin genes. These cDNA probes hybridized to calf thymus DNA to a slight, but significant, extent, and not at all to Micrococcus lysodektikus DNA. DNA complementary to Xenopus albumin mRNA did not cross-hybridize significantly with locust or chicken DNA. Further evidence for the evolutionary conservation of vitellogenin genes was obtained from Southern blot analysis of restriction endonuclease-digested genomic DNA from a variety of vertebrate and invertebrate oviparous animals (Xenopus, chicken, migratory and desert locusts, yellow meal worm, carab moth, and Mediterranean fruitfly). When probed with cloned vitellogenin cDNAs from Xenopus and migratory locust, the DNA of these organisms showed varying degrees of homology of parts of the vitellogenin coding sequences. Southern blot analysis also showed that a part of the sequence specified in the cloned Xenopus vitellogenin cDNA was represented as repetitive DNA in the locust genome. However, cloned locust vitellogenin cDNA hybridized to discrete fragments of the restricted vertebrate DNA. These studies demonstrate a remarkably high degree of conservation of insect, amphibian, and avian vitellogenin genes.