Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;...
Utility Patent
1999-08-31
2001-01-02
Patterson, Jr., Charles L. (Department: 1652)
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
C435S252300, C435S193000, C536S023200
Utility Patent
active
06168951
ABSTRACT:
FIELD OF THE INVENTION
This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding geranylgeranyl transferase subunits in plants and seeds.
BACKGROUND OF THE INVENTION
Lipids and proteins associate covalently to form lipid-linked proteins and noncovalently to form lipoproteins. The lipid portions of lipid-linked proteins anchor their attached proteins to membranes and mediate protein-protein interactions. Proteins form covalent attachments to lipids in several ways, one of which is the covalent attachment of isoprenoid groups, mainly the C
15
farnesyl and C
20
geranylgeranyl residues.
In mammals, geranylgeranyltransferase is known to catalyze the transfer of a geranyl-geranyl moiety from geranylgeranyl pyrophsophate to both cysteines in Rab proteins (Farnsworth, C. C. etal. (1994)
Proc Natl Acad Sci USA
91(25):11963-11967) Rab proteins are Ras-related small GTPases that are geranylgeranylated on cysteine residues located at or near their C termini. Mammalian protein geranylgeranyl transferases types 1 and 2 are heterodimers composed of an alpha and beta subunit. The alpha subunit shows homology to the alpha subunits of a closely related enzyme, farnesyltransferase.
Farnesyltransferases have been described in pea, tomato, and Arabidopsis, but have not been described in monocots. The plant farnesyltransferases also consist of alpha and beta subunits. The geranylgeranyl transferase beta subunit belongs to the protein prenyltransferase beta subunit family. The beta subunits of the type 1 and 2 geranylgeranyltransferases have not been previously described in plants. Work done in yeast has established that geranylgeranyltransferases are distinct from the closely related farnesyltransferases.
The mammalian geranylgeranyl transferases require the aid of a RAB escort protein (also called component A). RAB escort protein is required for Rab geranylgeranyl transferase activity in mammals. RAB escort protein binds unprenylated RAB proteins, presents it to the catalytic component B (alpha/beta subunit complex of geranylgeranyltransferase). RAB binding protein remains bound to the prenylated protein after the geranylgeranyl transfer reaction. Component A may be regenerated by transferring its prenylated RAB to a protein acceptor.
There is a great deal of interest in identifying the genes that encode geranylgeranyl transferase in plants. These genes may be used in plant cells to control cell growth. Accordingly, the availability of nucleic acid sequences encoding all or a portion of geranylgeranyl transferase proteins would facilitate studies to better understand cell growth in plants, provide genetic tools to enhance cell growth in tissue culture, increase the efficiency of gene transfer and help provide more stable transformations. Geranylgeranyl transferase proteins may also provide targets to facilitate design and/or identification of inhibitors of cell growth that may be useful as herbicides.
SUMMARY OF THE INVENTION
The instant invention relates to isolated nucleic acid fragments encoding geranylgeranyl transferase subunits. Specifically, this invention concerns an isolated nucleic acid fragment encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein and an isolated nucleic acid fragment that is substantially similar to an isolated nucleic acid fragment encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragment encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein.
An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of a geranylgeranyl transferase subunit selected from the group consisting of geranylgeranyl transferase type I beta subunit, type II beta subunit and Rab escort protein.
In another embodiment, the instant invention relates to a chimeric gene encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein, operably linked to suitable regulatory sequences, wherein expression of the chimeric gene results in production of levels of the encoded protein in a transformed host cell that is altered (i.e., increased or decreased) from the level produced in an untransformed host cell.
In a further embodiment, the instant invention concerns a transformed host cell comprising in its genome a chimeric gene encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein, operably linked to suitable regulatory sequences. Expression of the chimeric gene results in production of altered levels of the encoded protein in the transformed host cell. The transformed host cell can be of eukaryotic or prokaryotic origin, and include cells derived from higher plants and microorganisms. The invention also includes transformed plants that arise from transformed host cells of higher plants, and seeds derived from such transformed plants.
An additional embodiment of the instant invention concerns a method of altering the level of expression of a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein; and b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of altered levels of geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein in the transformed host cell.
An addition embodiment of the instant invention concerns a method for obtaining a nucleic acid fragment encoding all or a substantial portion of an amino acid sequence encoding a geranylgeranyl transferase type I beta subunit, type II beta subunit or Rab escort protein.
BRIEF DESCRIPTION OF THE SEQUENCE DESCRIPTIONS
The invention can be more fully understood from the following detailed description and the accompanying Sequence Listing which form a part of this application.
Table 1 lists the polypeptides that are described herein, the designation of the cDNA clones that comprise the nucleic acid fragments encoding polypeptides representing all or a substantial portion of these polypeptides, and the corresponding identifier (SEQ ID NO:) as used in the attached Sequence Listing. The sequence descriptions and Sequence Listing attached hereto comply with the rules governing nucleotide and/or amino acid sequence disclosures in patent applications as set forth in 37 C.F.R. §1.821-1.825.
TABLE 1
Geranylgeranyl Transferase Subunits
SEQ ID NO:
(Nucleo-
(Amino
Protein
Clone Designation
tide)
Acid)
Geranylgeranyl Transferase
cco1n.pk066.m2
1
2
Type II Beta Subunit
Geranylgeranyl Transferase
sf11.pk0074.b7
3
4
Type II Beta Subunit
Geranylgeranyl Transferase
w1su2.pk0001.h3
5
6
Type II Beta Subunit
Geranylgeranyl Transferase
sre.pk0040.h8
7
8
Type I Beta Subunit
Rab Escort Protein
r10n.pk0025.f10
9
10
Rab Escort Protein
wr1.pk0001.c3
11
12
The Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in
Nucleic Acids Res.
13:3021-3030 (1985) and in the
Biochemical J
219 (No. 2):345-373 (1984) which are herein incorporated by reference. The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. §1.822.
DETAILED DESCRIPTION OF THE INVENTION
In the context of this disclosure, a number of terms sh
Cahoon Rebecca E.
Kinney Anthony John
Rafalski J. Antoni
E. I. Du Pont de Nemours and Company
Patterson Jr. Charles L.
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