Distinct requirements for somatic and germline expression of a generally expressed Caernorhabditis elegans gene.
In screening for embryonic-lethal mutations in Caenorhabditis elegans, we defined an essential gene (let-858) that encodes a nuclear protein rich in acidic and basic residues. We have named this product nucampholin. Closely homologous sequences in yeast, plants, and mammals demonstrate strong evolutionary conservation in eukaryotes. Nucampholin resides in all nuclei of C. elegans and is essential in early development and in differentiating tissue. Antisense-mediated depletion of LET-858 activity in early embryos causes a lethal phenotype similar to characterized treatments blocking embryonic gene expression. Using transgene-rescue, we demonstrated the additional requirement for let-858 in the larval germline. The broad requirements allowed investigation of soma-germline differences in gene expression. When introduced into standard transgene arrays, let-858 (like many other C. elegans genes) functions well in soma but poorly in germline. We observed incremental silencing of simple let-858 arrays in the first few generations following transformation and hypothesized that silencing might reflect recognition of arrays as repetitive or heterochromatin-like. To give the transgene a more physiological context, we included an excess of random genomic fragments with the injected DNA. The resulting transgenes show robust expression in both germline and soma. Our results suggest the possibility of concerted mechanisms for silencing unwanted germiline expression of repetitive sequences.
Kelly
W G
WG
Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210, USA.
Xu
S
S
Montgomery
M K
MK
Fire
A
A
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins; D002875:Chromosomes; D005854:Germ Cells; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D016601:RNA-Binding Proteins; D017386:Sequence Homology, Amino Acid; D019076:Transgenes
D029742:Caenorhabditis elegans Proteins; C107534:LET-858 protein, C elegans; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins