Characterization of the translation products of the major mRNA species from rabbit lactating mammary glands and construction of bacterial recombinants containing casein and α-lactalbumin complementary DNA

Suard, Y.; Tosi, Mario; Kraehenbuhl, Jean–Pierre

doi:10.1042/bj2010081

Cited by 19 publications

(11 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4 and 5), a gene product of the liver (Morgan, 1969;MacKnight et al, 1980). However, in the rabbit, milk transferrin is synthesized by the mammary gland (Hauptle et al, 1979;Suard et al, 1982). Thus, in the rabbit, transferrin is expressed by two organs, the mammary gland and the liver.…”

Section: Discussionmentioning

confidence: 99%

“…In conclusion, our results indicate that rabbit caseins are made up of at least three structurally and antigenically distinct proteins that share similar properties with ruminant caseins. This is further supported by the immunoprecipitation of three individual peptides from cell-free translation of polyadenylated RNA isolated from lactating mammary gland [see the following paper (Suard et al, 1982)]. Antibodies have been raised against major whey proteins, including an iron-binding protein structurally identical with serum transferrin and a-lactalbumin.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk

Dayal¹,

Hürlimann²,

Suard³

et al. 1982

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

Caseins were separated from whey proteins by acid precipitation of skimmed rabbit milk. Whole casein was resolved by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis into three major bands with apparent relative molecular masses (Mr of 31 000, 29 000 and 25 000. On agarose/urea-gel electrophoresis whole casein gave three bands with electrophoretic mobilities alpha, beta and gamma. The three components were purified by DEAE-cellulose chromatography under denaturing and reducing conditions. Each was shown to have a different amino acid, hexose and phosphorus content, as well as non-identical peptide fragments after proteinase digestion. The 31 000 Da (dalton) protein, of alpha-electrophoretic mobility, had a high phosphorus content (4.38%, w/w); the 29 000 Da peptide, of gamma-mobility, had the highest hexose content (2.2%, w/w), contained 0.8 cysteine residue per 100 amino acid residues and was susceptible to chymosin digestion corresponding thus to kappa-casein; the 25 000 Da protein migrated to the beta-position. The rabbit casein complex is composed of at least three caseins, two of which (alpha- and kappa-caseins) are analogous to the caseins from ruminants. Although caseins are poor immunogens, specific antibodies were raised against total and purified polypeptides. The antiserum directed against whole casein recognized each polypeptide, each casein corresponding to a distinct precipitation line. The antisera directed against each casein polypeptide reacted exclusively with the corresponding casein and no antiserum cross-reaction occurred between the three polypeptides. From whey, several proteins were isolated, characterized and used as antigens to raise specific antibodies. An iron-binding protein with an apparent Mr of 80 000 was shown to be immunologically and structurally identical with serum transferrin.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk

Dayal¹,

Hürlimann²,

Suard³

et al. 1982

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…In such calculations, one does not take into account the amount of transferrin released from the basolateral membrane of the cells grown on floating gels. Since casein and a-lactalbumin probes are available (39), and transferrin cDNA has recently been constructed, it will now be possible to study the metabolism of the mRNAs coding for these milk proteins and to determine whether differences exist in the half-lives of the various mRNA populations.…”

Section: Hormonal Requirement For Milk Protein Cene Expressionmentioning

confidence: 99%

“…Total cellular RNA was denatured in 1 M glyoxal, 50% DMSO, 10 mM phosphate, pH 6.5, 0.1% SDS for 5 min at 50°C (following a modification of the procedure of McMaster and Carmichael [28]) and was clectrophoresed on a horizontal agarose slab gel in 10 mM phosphate buffer, pH 6.5. The RNA was transferred onto nitrocellulose filters following the protocol of Thomas (42) and was hybridized to cloned 32p-labeled ,/-casein cDNA (39). Hybridization with polyadenylated RNA extracted from lactating mammary gland served as a control.…”

Section: Dna Synthesis and Cell Countsmentioning

confidence: 99%

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Ym¹,

Mt²,

Farinon³

et al. 1983

The Journal of Cell Biology

View full text Add to dashboard Cite

We analyzed the synthesis of DNA, the rate of cell proliferation, and the expression of milk protein genes in mammary cells grown as primary cultures on or in collagen gels in chemically defined media. We assessed DNA synthesis and cell growth, measured by [aH]-thymidine incorporation into acid-insoluble material, DNA content, and cell counts, in a progesterone-and prolactin-containing medium. In some experiments, cultures were pulsed for I h with [3H]thymidine and dissociated into individual cells which were cytocentrifuged and processed for immunocytochemistry and autoradiography. We analyzed expression of milk protein genes at the transcriptional, translational, and posttranslational levels in a progesteronedepleted medium in the presence or absence of prolactin. We measured protein secretion by radioimmunoassays with antisera directed against caseins, ~-lactalbumin and milk transferrin. We determined protein synthesis by incorporating radio-labeled amino acids into acid-precipitable material and by immunoprecipitating biosynthetically labeled milk proteins. We assessed the accumulation of casein mRNA by hybridizing total cellular RNA extracted from cultured cells with 32p-labeled casein cDNA probes. On attached collagen gels, the cells synthesized DNA and replicated until they became confluent. The overall protein synthetic activity was low, and no milk proteins were synthesized or secreted even in the presence of prolactin. The block in milk protein gene expression was not restricted to translational or posttranslational events but also included transcription, since no casein mRNA accumulated in these cells. On floating gels, protein synthesis was threefold higher than in cells from attached gels. Overall protein synthesis as well as casein and ~-lactalbumin synthesis and secretion were prolactindependent with maximal stimulation at around 10 -9 M. A marked inhibition occurred at higher hormone concentrations. Casein mRNA accumulated in these cells, provided prolactin was present in the medium. In contrast, these cells did not synthesize DNA, nor did they replicate. In embedding gels, the rate of cell proliferation was exponential over 25 d with a doubling time of ~70 h. The overall protein synthesis increase was parallel in time with the increase in cell number. Caseins and ~-lactalbumin (in contrast to transferrin) were synthesized only in the presence of prolactin. We observed the same hormone dependency as with cells growing on floating gels. The number of casein-and transferrin-positive cells was measured after dissociating the cell cultures. At day 12, 60% of the total cells stored transferrin in small cytoplasmic vesicles, whereas only 25% of the cells accumulated casein. Differences in the organization and in the shape of mammary cells depending on cell surface conditions suggest that the geometry of the cells, their interaction with extracellular matrix constitutents, and cellto-cell interactions play a role in the expression of two mammary functions: DNA synthesis and growth, as well as m...

show abstract

“…The probes were two recombinant plasmids kindly provided by Dr Devinoy. who isolated them from a rabbit mammary gland cDNA library constructed by Suard et al (1982). The two probes, a casein (928) and |S casein (924).…”

Section: Methodsmentioning

confidence: 99%

Localization of the α and β casein genes to the q24 region of chromosome 12 in the rabbit (<i>Oryctolagus cuniculu</i><i>s</i> L.) by in situ hybridization

Gellin¹,

Echard²,

Yerle³

et al. 1985

Cytogenet Genome Res

View full text Add to dashboard Cite

The syntenic α and β casein genes were localized in the rabbit by chromosomal in situ hybridization, using a mixture of two radioactive cDNA probes corresponding to these two genes. Highly significant labeling was observed on chromosome 12. A total of 175 silver grains was found on chromosomes in the 193 mitoses studied; 18% of the grains were on chromosome 12, and 42% of the grains on this chromosome were localized to the 12q24 region. Statistical analysis revealed that this labeling was highly significant.

show abstract

Characterization of the translation products of the major mRNA species from rabbit lactating mammary glands and construction of bacterial recombinants containing casein and α-lactalbumin complementary DNA

Cited by 19 publications

References 54 publications

Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk

Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Localization of the α and β casein genes to the q24 region of chromosome 12 in the rabbit (<i>Oryctolagus cuniculu</i><i>s</i> L.) by in situ hybridization

Contact Info

Product

Resources

About