Dominant positive hnRNP-E1 polypeptide compositions and methods
View Patent ↗Described herein are compositions relating to engineered hnRNP-E1 variant polypeptides, nucleic acids encoding such polypeptides, engineered hnRNP-E1 compositions, and methods of use thereof. In some embodiments, the engineered hnRNP-E1 polypeptide contains a C293S substitution and retains the ability to bind to a poly(rC)- and poly(U)-rich 5′-UTR element in its cognate mRNA targets in the absence of homocysteine. In some cases, the engineered hnRNP-E1 compositions provided herein are useful to increase the translation of a subset of mRNAs or to treat certain health conditions as described herein.
1. An isolated variant of a native hnRNP-E1 polypeptide, wherein the variant hnRNP-E1 polypeptide comprises (i) at least one amino acid substitution at an amino acid residue selected from C293, C54, C158, and C201 of SEQ ID NO:1, wherein the amino acid substitution replaces a native cysteine residues with a serine residue and (ii) an RNA binding portion that binds a single-stranded RNA that comprises SEQ ID NO:6 in the absence of homocysteine and is optionally linked to a protein transduction domain.
2. The variant hnRNP-E1 polypeptide of claim 1 , wherein the variant hnRNP-E1 polypeptide binds the single-stranded RNA comprising SEQ ID NO:6 independent of homocysteine concentration.
3. The variant hnRNP-E1 polypeptide of claim 1 , wherein the variant hnRNP-E1 polypeptide comprises an amino acid sequence at least 95% identical to any one of SEQ ID NOs:2-5.
4. The variant hnRNP-E1 polypeptide of claim 1 , wherein the variant hnRNP-E1 polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:2-5.
5. The variant hnRNP-E1 polypeptide of claim 4 , wherein the amino acid sequence is SEQ ID NO:2.
6. The variant hnRNP-E1 polypeptide of claim 1 , wherein the is linked to the protein transduction domain.
7. The variant hnRNP-E1 polypeptide of claim 6 , wherein the protein transduction domain comprises any of SEQ ID NOs:7-11.
8. A pharmaceutical composition comprising the variant hnRNP-E1 polypeptide of claim 1 and a pharmaceutically acceptable carrier.
9. A method for increasing translation of a target mRNA in a mammalian cell, the method comprising the step of:
providing in the mammalian cell the polypeptide of claim 1 , wherein the target mRNA comprises a poly(rC)- and poly(U)-rich 5′ UTR, thereby increasing translation of the target mRNA in the mammalian cell.
10. The method of claim 9 , wherein the polypeptide comprises an RNA binding portion comprising SEQ ID NO: 2.
11. The method of claim 9 , wherein the providing step is selected from the group consisting of administering the polypeptide to the cell and expressing the polypeptide within the cell.
12. The method of claim 11 , wherein the expressing step includes delivering into the mammalian cell a nucleic acid encoding the polypeptide whereby the polypeptide is expressed.
13. The method of claim 9 , wherein the target mRNA is selected from the group consisting of folate receptor mRNA, hnRNP-E1 mRNA, collagen alpha (I) mRNA, human papillomavirus type 16 L2 mRNA, human herpesvirus 8 mRNA, erythropoietin mRNA, human alpha-globin mRNA, human beta-globin mRNA, μ-opioid receptor mRNA, androgen receptor mRNA, p21waf mRNA, tyrosine hydroxylase mRNA, and neurofilament M mRNA.