Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1998-05-27
2001-08-14
Hauda, Karen M. (Department: 1632)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C536S023100, C536S023500, C435S320100, C435S325000
Reexamination Certificate
active
06274338
ABSTRACT:
BACKGROUND OF THE INVENTION
The Maf family of proteins are a sub-family of AP-1/CREB/ATF proteins. The first member of the family to be identified, the v-maf oncogene, was originally isolated from a spontaneous musculoaponeurotic fibrosarcoma of chicken and identified as the transforming gene of the avian retrovirus, AS42 (Nishizawa, M. et al. (1989)
Proc. Natl. Acad. Sci. USA
86:7711-7715). V-maf encodes a 42 kd basic region/leucine zipper (b-zip) protein with homology to the c-fos and c-jun oncogenes. Its cellular homologue, the c-maf proto-oncogene, which has been isolated from murine cells, has only two structural changes in the coding region from &ngr;-maf(Kataoka, K. et al. (1993)
J. Virol
. 67:2133-2141). The maf family includes c-Maf, mafb, a human retina-specific protein Nrl (Swaroop, A. et al. (1992)
Proc. Natl. Acad. Sci. USA
89:266-270), mafk, mafF, mafG and p18. The latter four, mafK, mafF, mafG and p18, each encode proteins that lack the amino terminal two thirds of c-Maf that contains the transactivating domain (“small maf proteins”) (Fujiwara, K. T. et al. (1993)
Oncogene
8:2371-2380; Igarashi, K. et al. (1995)
J. Biol. Chem
. 270:7615-7624; Andrews, N. C. et al. (1993)
Proc. Natl. Acad. Sci. USA
90:11488-11492; Kataoka, K. et al. (1995)
Mol. Cell. Biol
. 15:2180-2190).
C-Maf and other Maf family members form homodimers and heterodimers with each other and with Fos and Jun, consistent with the known ability of the AP-1 proteins to pair with each other (Kerppola, T. K. and Curran, T. (1994)
Oncogene
9:675-684; Kataoka, K. et al. (1994)
Mol. Cell. Biol
. 14:700-712). The DNA target sequence to which c-Maf homodimers bind, termed the c-Maf response element (MARE), is a 13 or 14 bp element which contains a core TRE (T-MARE) or CRE (C-MARE) palindrome respectively, c-Maf has been shown to stimulate transcription from the Purkinje neuron-specific promoter L7 (Kurscher, C. and Morgan, J. I. (1994)
Mol. Cell. Biol
. 15:246-254) and Nrl has been shown to drive expression of the QR1 retina-specific gene (Swaroop, A. et al. (1992)
Proc. Natl. Acad. Sci. USA
89:266-270). Additionally, the small mafs have been shown to function as repressors of &agr; and &bgr;-globin transcription when bound as homodimers but are essential as heterodimeric partners with the erythroid-specific factor p45NF-E2 to activate globin gene transcription (Kataoka, K. et al. (1995)
Mol. Cell. Biol
. 15:2180-2190; Igarashi, K. et al. (1994)
Nature
367:568-572). MafK overexpression has been shown to induce erythroleukemia cell differentiation (Igarashi, K. et al. (1995)
Proc. Natl. Acad. Sci. USA
92:7445-7449). Moreover, c-Maf has been shown to control the tissue-specific expression of the cytokine interleukin-4 in T helper 2 (Th2) cells (Ho, I-C. et al. (1996)
Cell
85:973-983).
The nucleotide sequence of the mouse c-maf proto-oncogene, and predicted amino acid sequence for the mouse c-Maf protein, have been described (Kurscher, C. and Morgan, J. I. (1995)
Mol. Cell. Biol
. 15:246-254; and Genbank Accession number S74567). The nucleotide sequence of the chicken c-maf proto-oncogene, and predicted amino acid sequence for the chicken c-Maf protein, also have been described (Kataoka et al., Genbank Accession number D28596). However, these non-human c-Maf compositions may not function optimally in human cells and, moreover, use of these compositions in humans is likely to stimulate an immune response, since the chicken or mouse c-Maf would be recognized as “foreign” by the human immune system. Accordingly, there is still a need for human c-Maf compositions that are suitable for use in humans.
SUMMARY OF THE INVENTION
This invention provides human c-Maf compositions. In particular, this invention provides isolated nucleic acid molecules encoding human c-Maf and isolated human c-Maf protein. Since the c-Maf compositions of the invention are human-derived, they function optimally in human cells (compared with non-human c-Maf compositions) and do not stimulate an immune response in humans.
One aspect of the invention pertains to an isolated nucleic acid molecule comprising a nucleotide sequence encoding human c-Maf. In a preferred embodiment, the nucleic acid molecule comprises the nucleotide sequence of the coding region of the NheI/XbaI insert of plasmid pHu-c-Maf (ATCC Accession No. 98671). In another preferred embodiment, the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 1. In other embodiments, the nucleic acid molecule has at least 98% nucleotide identity, more preferably 99% nucleotide identity, and even more preferably 99.5% nucleotide identity with the nucleotide sequence of SEQ ID NO: 1 or the nucleotide sequence of the NheI/XbaI insert of plasmid pHu-c-Maf (ATCC Accession No. 98671). In yet another embodiment, the nucleic acid molecule comprises the nucleotide sequence of the NheI/XbaI insert of plasmid pHu-c-Maf (ATCC Accession No. 98671).
The isolated nucleic acid molecules of the invention encoding human c-Maf can be incorporated into a vector, such as an expression vector, and this vector can be introduced into a host cell. The invention also provides a method for producing a human c-Maf protein by culturing a host cell of the invention (carrying a hu-c-Maf expression vector) in a suitable medium until a human c-Maf protein is produced. The method can further involve isolating the human c-Maf protein from the medium or the host cell.
Another aspect of the invention pertains to an isolated human c-Maf protein. Preferably, the human c-Maf protein comprises the amino acid sequence encoded by the coding region of the NheI/XbaI insert of plasmid pHu-c-Maf (ATCC Accession No. 98671). In another preferred embodiment, the protein comprises the amino acid sequence of SEQ ID NO: 2. In other embodiments, the protein has at least 98% amino acid identity, more preferably 99% amino identity, and even more preferably 99.5% amino acid identity with SEQ ID NO: 2 or the protein encoded by the coding region of the NheI/XbaI insert of plasmid pHu-c-Maf (ATCC Accession No. 98671).
Fusion proteins, comprising a human c-Maf protein operatively linked to a polypeptide other than human c-Maf, are also encompassed by the invention, as well as antibodies that specifically bind a human c-Maf protein. The antibodies can be, for example, polyclonal antibodies or monoclonal antibodies. In one embodiment, the antibodies are coupled to a detectable substance.
Another aspect of the invention pertains to a nonhuman transgenic animal that contains cells carrying a transgene encoding a human c-Maf protein.
Yet another aspect of the invention pertains to a method for detecting the presence of human c-Maf in a biological sample. The method involves contacting the biological sample with an agent capable of detecting an indicator of human c-Maf activity such that the presence of human c-Maf is detected in the biological sample. The invention also provides a method for modulating human c-Maf activity in a cell comprising, involving contacting the cell with an agent that modulates human c-Maf activity such that human c-Maf activity in the cell is modulated.
Still another aspect of the invention pertains to methods for identifying a compound that modulates the activity of a human c-Maf protein. These methods generally involve:
providing an indicator composition that comprises a human c-Maf protein;
contacting the indicator composition with a test compound; and
determining the effect of the test compound on the activity of the human c-Maf protein in the indicator composition to thereby identify a compound that modulates the activity of a human c-Maf protein. In a preferred embodiment, the indicator composition comprises a human c-Maf protein and a DNA molecule to which the human c-Maf protein binds and the effect of the test compound on the activity of the human c-Maf protein is determined by evaluating the binding of the human c-Maf protein to the DNA molecule in the presence and absence of the test compound. In another preferred embodiment, the indicator composition is a cell comprising a
Douhan, III John
Glimcher Laurie H.
DeConti, Jr. Giulio A.
Hauda Karen M.
Lahive & Cockfield LLP
President and Fellows of Harvard College
Woitach Joseph
LandOfFree
Human c-Maf compositions and methods of use thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Human c-Maf compositions and methods of use thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Human c-Maf compositions and methods of use thereof will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2550685