Targeting of E. coli beta-galactosidase to the nucleus in yeast
- PMID: 6323016
- DOI: 10.1016/0092-8674(84)90055-2
Targeting of E. coli beta-galactosidase to the nucleus in yeast
Abstract
In order to identify determinants governing nuclear protein localization, we constructed a set of hybrid genes by fusing the S. cerevisiae gene, MAT alpha 2, coding for a presumptive nuclear protein, and the E. coli gene, lacZ, coding for beta-galactosidase. The resultant hybrid proteins contain 3, 13, 25, 67, or all 210 amino acids of wild-type alpha 2 protein at the amino terminus and a constant, enzymatically active portion of beta-galactosidase at the carboxy terminus. Indirect immunofluorescence and subcellular fractionation studies with yeast cells containing the alpha 2-LacZ hybrid proteins indicate that the alpha 2 segment can direct localization of beta-galactosidase to the nucleus. A segment as small as 13 amino acids from alpha 2 is sufficient for this localization. Comparison of amino acid sequences of other nuclear proteins with this region of alpha 2 reveals a sequence that may be necessary for nuclear targeting. Production of some alpha 2-LacZ hybrid proteins causes cell death, perhaps as a result of improper or incomplete localization. These studies also indicate that the alpha 2 protein, argued on genetic grounds to be a negative regulator, acts in the yeast nucleus.
Similar articles
-
Intracellular targeting and import of an F1-ATPase beta-subunit-beta-galactosidase hybrid protein into yeast mitochondria.Proc Natl Acad Sci U S A. 1984 Jul;81(13):3983-7. doi: 10.1073/pnas.81.13.3983. Proc Natl Acad Sci U S A. 1984. PMID: 6330727 Free PMC article.
-
Expression of Ty-lacZ fusions in Saccharomyces cerevisiae.Nucleic Acids Res. 1984 Feb 10;12(3):1627-40. doi: 10.1093/nar/12.3.1627. Nucleic Acids Res. 1984. PMID: 6322112 Free PMC article.
-
Construction and use of gene fusions to lacZ (beta-galactosidase) that are expressed in yeast.Methods Enzymol. 1983;101:167-80. doi: 10.1016/0076-6879(83)01012-5. Methods Enzymol. 1983. PMID: 6310320 No abstract available.
-
New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase.Gene. 1983 Nov;25(1):71-82. doi: 10.1016/0378-1119(83)90169-5. Gene. 1983. PMID: 6319233
-
Genetic studies on protein export in bacteria.Curr Top Microbiol Immunol. 1986;125:5-27. doi: 10.1007/978-3-642-71251-7_2. Curr Top Microbiol Immunol. 1986. PMID: 3091324 Review. No abstract available.
Cited by
-
GAL2 codes for a membrane-bound subunit of the galactose permease in Saccharomyces cerevisiae.J Bacteriol. 1986 Apr;166(1):313-8. doi: 10.1128/jb.166.1.313-318.1986. J Bacteriol. 1986. PMID: 3082856 Free PMC article.
-
Monoclonal antibody to a protein of the nucleus and mitotic spindle of mammalian cells. Localization and synthesis throughout the cell cycle.Chromosoma. 1985;92(4):297-303. doi: 10.1007/BF00329813. Chromosoma. 1985. PMID: 3899548
-
Identification of domains involved in nuclear uptake and histone binding of protein N1 of Xenopus laevis.EMBO J. 1988 Jun;7(6):1605-14. doi: 10.1002/j.1460-2075.1988.tb02986.x. EMBO J. 1988. PMID: 3168999 Free PMC article.
-
A method for constructing single-copy lac fusions in Salmonella typhimurium and its application to the hemA-prfA operon.J Bacteriol. 1992 Jan;174(1):245-53. doi: 10.1128/jb.174.1.245-253.1992. J Bacteriol. 1992. PMID: 1309519 Free PMC article.
-
Separate basic region motifs within the adeno-associated virus capsid proteins are essential for infectivity and assembly.J Virol. 2006 Jun;80(11):5199-210. doi: 10.1128/JVI.02723-05. J Virol. 2006. PMID: 16699000 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Molecular Biology Databases
