The mouse p97 (CDC48) gene. Genomic structure, definition of transcriptional regulatory sequences, gene expression, and characterization of a pseudogene.
Here we present the first description of the genomic organization, transcriptional regulatory sequences, and adult and embryonic gene expression for the mouse p97(CDC48) AAA ATPase. Clones representing two distinct p97 genes were isolated in a genomic library screen, one of them likely representing a non-functional processed pseudogene. The coding region of the gene encoding the functional mRNA is interrupted by 16 introns and encompasses 20.4 kilobase pairs. Definition of the transcriptional initiation site and sequence analysis showed that the gene contains a TATA-less, GC-rich promoter region with an initiator element spanning the transcription start site. Cis-acting elements necessary for basal transcription activity reside within 410 base pairs of the flanking region as determined by transient transfection assays. In immunohistological analyses, p97 was widely expressed in embryos and adults, but protein levels were tightly controlled in a cell type- and cell differentiation-dependent manner. A remarkable heterogeneity in p97 immunostaining was found on a cellular level within a given tissue, and protein amounts in the cytoplasm and nucleus varied widely, suggesting a highly regulated and intermittent function for p97. This study provides the basis for a detailed analysis of the complex regulation of p97 and the reagents required for assessing its functional significance using targeted gene manipulation in the mouse.
Müller
J M
JM
Cell Biology Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London WC2A 3PX, UK.
Meyer
H H
HH
Ruhrberg
C
C
Stamp
G W
GW
Warren
G
G
Shima
D T
DT
D000251:Adenosine Triphosphatases; D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000502:physiology*; D002454:Cell Differentiation; D002455:Cell Division; D004331:Drosophila melanogaster; D005786:Gene Expression Regulation; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011544:Pseudogenes; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic; D000074405:Valosin Containing Protein; D014982:Xenopus laevis
D018797:Cell Cycle Proteins; D000251:Adenosine Triphosphatases; D000074405:Valosin Containing Protein