Identification and characterization of functional genes encoding the mouse major urinary proteins.

Held, William A.; Gallagher, John F.; Hohman, Colleen; Kuhn, Nancy J.; Sampsell, Bonnie M.; Hughes, Robert G.

doi:10.1128/mcb.7.10.3705

Cited by 32 publications

(36 citation statements)

References 34 publications

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“…Expression of the hybrid gene was studied by probing Northern blots with probes specific for HSV tk and by assaying HSV thymidine kinase activity in tissue homogenates. Group 1 MUP mRNA is present in male and female liver and also in small amounts in the prelactation mammary gland, but not in other tissues (25,50). The foreign HSV tk mRNA and thymidine kinase activity, however, were consistently present not only in male and female liver but also in testis and in preputial glands.…”

Section: Resultsmentioning

confidence: 93%

See 1 more Smart Citation

A Mup promoter-thymidine kinase reporter gene shows relaxed tissue-specific expression and confers male sterility upon transgenic mice.

Al‐Shawi¹,

Burke²,

Jones³

et al. 1988

Mol. Cell. Biol.

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A hybrid gene was made by fusing the 2.2-kilobase 5' promoter region of a mouse group 1 major urinary protein (Mup) gene to the coding region of the herpes simplex virus type 1 thymidine kinase gene (HSV tk) and introduced into the genomes of mice by microinjection. Transgenic Go males were sterile, or when fertile did not transmit the foreign gene, and the transgenic male descendants of Go females were also sterile. Seven "lines" were established by breeding from Go females and their transgenic female descendants. Six lines expressed HSV thymidine kinase activity in the liver, and activity correlated perfectly with the presence of HSV tk RNA. In three of four lines examined, expression was lower in female than in male liver, and in these lines the same sex difference was observed in the rate of run-on transcription of the foreign genes in liver nuclei. When females of one of the sexually dimorphic lines were treated with testosterone, the levels of HSV tk RNA and thymidine kinase activity were increased, although not to male levels. In these aspects of liver expression, and also in a lack of expression in seven other tissues, the hybrid gene exhibits many of the characteristics of an endogenous group 1 Mup gene. However, the gene was also expressed (at high levels) in the preputial gland and testis, two tissues in which Mup genes are not expressed. The gene, when introduced into five of the seven lines, carried a copy of the Escherichia coli supF gene attached beyond the 3' end of the HSV tk gene, but this did not affect the overall expression pattern. All of the lines were male sterile and expressed HSV thymidine kinase in the testis, but one line showed no activity in the liver, and another showed none in the preputial gland. Testicular expression is therefore the likely cause of sterility. Data are described which suggest that the causes of misexpression in the preputial gland and testis are different. Since expression in each tissue occurred in several lines, the structure of the hybrid gene must be responsible in each case. Five intensively studied lines showed at least four consistently different patterns of relative expression in preputial gland, testis, male liver, and female liver. These differences do not correlate in any way with the copy number of the foreign gene in the different lines and must be due to some other aspect of line-specific integration.The mouse major urinary proteins (MUPs) are coded for by a family of about 35 genes clustered on chromosome 4 (3, 12, 23). Four groups of Mup genes have been identified, the largest of which are the group 1 genes and the probably inert group 2 pseudogenes (3, 12, 25, 35, 51; R. Al-Shawi, P. Ghazal, A. J. Clark, and J. 0. Bishop, submitted for publication). Most group 1 genes are expressed exclusively in the liver, while a small subset are also expressed in the mammary glands of pregnant females (50, 52). Of the two smaller groups, the group 3 genes are also expressed in the liver, but under hormonal control different from the group 1 genes (35), and...

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Section: Resultsmentioning

confidence: 93%

“…Four groups of Mup genes have been identified, the largest of which are the group 1 genes and the probably inert group 2 pseudogenes (3,12,25,35,51; R. Al-Shawi, P. Ghazal, A. J. Clark, and J. 0.…”

mentioning

confidence: 99%

A Mup promoter-thymidine kinase reporter gene shows relaxed tissue-specific expression and confers male sterility upon transgenic mice.

Al‐Shawi¹,

Burke²,

Jones³

et al. 1988

Mol. Cell. Biol.

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“…With the exception of a localized AT-rich hypervariable region (16; Al-Shawi et al, in press), the 5'-flanking regions of group 1 Mup genes are highly conserved (12,16). Pairwise differences are about 2% in 800 base pairs of 5'-flanking DNA (16).…”

Section: Resultsmentioning

confidence: 99%

“…Pairwise differences are about 2% in 800 base pairs of 5'-flanking DNA (16). It will be interesting to determine whether one or more of these minor differences is responsible for the large difference between BLI and BSJ expression in female liver.…”

Section: Resultsmentioning

confidence: 99%

Differential expression in male and female mouse liver of very similar mRNAs specified by two group 1 major urinary protein genes.

McIntosh

Bishop

1989

Mol. Cell. Biol.

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The major urinary proteins of the mouse are encoded by a large multigene family composed of several distinct groups of genes distinguished by differences in sequence and expression characteristics. The genes in the largest group (group 1) show greater than 99% pairwise similarity in their exons. By hybridization between RNA and a specifically designed oligonucleotide, we confirmed that genes of this group are expressed mainly in the liver. By using additional gene-specific oligonucleotide probes, we have been able to distinguish between the species of mRNA corresponding to two of these genes and to measure their abundance in male and female liver. Both mRNAs are present in male liver at high but different levels. Both are also present in female liver, one at a much lower level than in the male and the second at a very low level indeed. Both are present at male levels in the livers of females induced with testosterone. These results show unequivocally that the expression of different group 1 Mup genes is differentially influenced by the hormonal status of the mouse.

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“…As a consequence, nonsense codon recognition within env mRNA would be too late to affect nuclear RNA metabolism. Since MUP has been shown to be secreted when produced from either pMUP in mouse L cells (25) or pCMV-MUP in a variety of cell types (53), CMV-MUP mRNA should become associated with the endoplasmic reticulum after translation initiates. Whether or not this is concomitant with or subsequent to translocation through the nuclear pore remains speculative for any mRNA.…”

Section: Discussionmentioning

confidence: 99%

Nonsense but not missense mutations can decrease the abundance of nuclear mRNA for the mouse major urinary protein, while both types of mutations can facilitate exon skipping.

1994

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In an effort to understand the mechanisms by which nonsense codons affect RNA metabolism in mammalian cells, nonsense mutations were generated within the gene for the secretory major urinary protein (MUP) of mice.The translation of MUP mRNA normally begins within exon 1 and terminates within exon 6, the penultimate exon.Through the use of Northern (RNA) blot hybridization and assays that couple reverse transcription and PCR, a nonsense mutation within codon 50 of exon 2 or codon 143 of exon 5 was found to reduce the abundance of fully spliced, nuclear MUP mRNA to 10 to 201% of normal without an additional reduction in the abundance of cytoplasmic mRNA. In contrast, a nonsense mutation within codon 172 of exon 5 was found to have no effect on the abundance of MUP mRNA. These findings suggest that a boundary between nonsense mutations that do and do not reduce the abundance of nuclear mRNA exists within the exon preceding the exon that harbors the normal site of translation termination. In this way, the boundary is analogous to the boundary that exists within the penultimate exon of the human gene for the cytosolic enzyme triosephosphate isomerase. Assays for exon skipping, i.e., the removal of an exon as a part of the flanking introns during the process of splicing, reveal that 0.1, 2.0, and 0.1% of MUP mRNA normally lack exon 5, exon 6, and exons 5 plus 6, respectively. Relative to normal, the two nonsense mutations within exon 5 increase the abundance of RNA lacking exon 5 on average 20-fold and increase the abundance of RNA lacking exons 5 plus 6 on average 5-fold. Since only one of these nonsense mutations also reduces the abundance of fully spliced nuclear mRNA to 10 to 20%o of normal, the two mechanisms by which a nonsense mutation can alter nuclear RNA metabolism must be distinct. The analysis of missense mutations within codons 143 and 172, some of which retain the nonsense mutation, indicates that the reduction in the abundance of fully spliced nuclear mRNA is dependent upon the premature termination of MUP mRNA translation, whereas skipping is attributable to nonsense mutation-mediated changes in exon 5 structure rather than to the premature termination of translation. The increase in exon 5 skipping by either the nonsense or missense mutations within codon 172 correlates with a decrease in the complementarity of exon 5 to Ul snRNA. This suggests that a 5' splice site may extend as far as 12 nucleotides into the upstream exon, which is, to our knowledge, the largest extension.In eukaryotes, protein-encoding genes that harbor a nonsense codon because of a nonsense or frameshift mutation often produce RNA that is metabolized abnormally. For some genes, the product mRNA is increased in abundance because the premature termination of translation precludes ribosome translocation either across a specific sequence in the coding region, as exemplified by the c-fos, c-myc, and MA Tot] mRNAs (22,49,57,58,69), or sufficiently close to a higher-order structure in the 3' untranslated region, as exemplified by specifi...

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Identification and characterization of functional genes encoding the mouse major urinary proteins.

Cited by 32 publications

References 34 publications

A Mup promoter-thymidine kinase reporter gene shows relaxed tissue-specific expression and confers male sterility upon transgenic mice.

A Mup promoter-thymidine kinase reporter gene shows relaxed tissue-specific expression and confers male sterility upon transgenic mice.

Differential expression in male and female mouse liver of very similar mRNAs specified by two group 1 major urinary protein genes.

Nonsense but not missense mutations can decrease the abundance of nuclear mRNA for the mouse major urinary protein, while both types of mutations can facilitate exon skipping.

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