Chromatin associated proteins

Details Chromatin associated proteins Chromatin associated proteins

1999-06-29

2001-10-23

Prouty, Rebecca E. (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Oxidoreductase

C435S069100, C435S183000, C435S419000, C435S410000, C435S006120, C435S252300, C435S320100, C435S254200, C536S023100, C536S023200, C530S350000, C800S278000, C800S295000

Reexamination Certificate

active

06306632

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 chromatin associated proteins in plants and seeds.
BACKGROUND OF THE INVENTION
The human Lamin B receptor (LBR) belongs to the ERG4/ERG24 family of nuclear envelope inner membrane proteins. It anchors the lamina and the heterochromatin to the inner nuclear membrane. LBR can interact with chromodomain proteins. LBR has an amino-terminal domain of approximately 200 amino acids followed by a carboxyl-terminal domain that is similar in sequence to yeast and plant sterol reductases. Two LBR-like genes have recently been identified in humans which have strong carboxyl-terminal domains of LBR and sterol reductases (Pezhman et al. (1998)
Genomics
54(3):469-476). The human LBR/sterol reductase like proteins are localized to the endoplasmic reticulum. These LBR/sterol reductase proteins may define a human gene family encoding proteins of the inner nuclear membrane and endoplasmic reticulum that function in nuclear organization and/or sterol metabolism.
In the nucleus LBR undergoes phosphorylation by CDC2 protein kinase in mitosis when the inner nuclear membrane breaks down into vesicles that dissociate from the lamina and the chromatin. It is phosphorylated by different protein kinases in interphase when the membrane is associated with these structures. Phosphorylation of LBR proteins may be responsible for some of the alternations in chromatin organization and nuclear structure which occur at various times during the cell cycle. To date, a Lamin B receptor has not been identified in plants. A plant Lamin B receptor could be used to manipulate cell cycle regulation and plant transformability.
Accordingly, the availability of nucleic acid sequences encoding all or a portion of these proteins would facilitate studies to better understand transcritional regulation, cell cycle progression, and developmental events in eucaryotic cells. It would also provide genetic tools for the manipulation of cell cycle regulation and increase the efficiency of transformation.
SUMMARY OF THE INVENTION
The instant invention relates to isolated nucleic acid fragments encoding chromatin associated proteins. Specifically, this invention concerns an isolated nucleic acid fragment encoding a lamin B receptor/sterol reductase and an isolated nucleic acid fragment that is substantially similar to an isolated nucleic acid fragment encoding a lamin B receptor/sterol reductase. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragment encoding lamin B receptor/sterol reductase.
An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of a lamin B receptor/sterol reductase.
In another embodiment, the instant invention relates to a chimeric gene encoding a lamin B receptor/sterol reductase, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding a lamin B receptor/sterol reductase, 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 lamin B receptor/sterol reductase, 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 lamin B receptor/sterol reductase in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a lamin B receptor/sterol reductase; 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 lamin B receptor/sterol reductase 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 lamin B receptor/sterol reductase.
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
Chromatin Associated Proteins
SEQ ID NO:
Clone
(Amino
Protein
Designation
(Nucleotide)
Acid)
Lamin B/Sterol Reductase
bms1.pk0009.e4
1
2
(corn)
Lamin B/Sterol Reductase
rlr2.pk0031.g9
3
4
(rice)
Lamin B/Sterol Reductase
wr1.pk0039.d9
5
6
(wheat)
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 Research
13:3021-3030 (1985) and in the
Biochemical Journal
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 shall be utilized. As used herein, a “nucleic acid fragment” is a polymer of RNA or DNA that is single- or double-stranded, optionally containing synthetic, non-natural or altered nucleotide bases. A nucleic acid fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA.
As used herein, “substantially similar” refers to nucleic acid fragments wherein changes in one or more nucleotide bases results in substitution of one or more amino acids, but do not affect the functional properties of the polypeptide encoded by the nucleotide sequence. “Substantially similar” also refers to nucleic acid fragments wherein changes in one or more nucleotide bases does not affect the ability of the nucleic acid fragment to mediate alteration of gene expression by gene silencing through for example antisense or co-suppression technology. “Substantially similar” also refers to modifications of the nucleic acid fragments of the instant invention such as deletion or insertion of one or more nucleotides that do not substantially affect the functional properties of the resulting transcript vis-à-vis the ability to mediate gene silencing or alteration of the functional properties of the resulting protein molecule. It is therefore understood that the invention encompasses more than the specific exemplary nucleotide or amino acid sequences and includes functional equivalents thereof.
For example, it is well known in the art that antisense suppression and co-suppression of gene expression may be accomplished using nucleic acid fragments representing less than the entire coding region of a gene, and by nucleic acid fragments that do not share 100% sequence identity with the gene to be suppressed. Moreover, altera

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