Kyushu University

Hiroshima University

Yusuke Yagi

Takahiro Nakamura

Keiko Kobayashi

Takashi Yamamoto

Tetsushi Sakuma

Yasuyuki Okawa

A method for designing a protein capable of binding in an RNA base selective manner or RNA base sequence specific manner is provided. The protein of the present invention is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1 (wherein Helix A is a moiety of 12-amino acid length capable of forming an &#x3b1;-helix structure, and is represented by the formula 2, wherein, in the formula 2, Ato Aindependently represent an amino acid; X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an &#x3b1;-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3, wherein, in the formula 3, the amino acids are numbered 'i' (&#x2212;1), 'ii' (&#x2212;2), and so on from the C-terminus side, provided that Lto Lmay not exist), and combination of three amino acids A, Aand L, or combination of two amino acids A, and Lis a combination corresponding to a target RNA base or base sequence.

Method for designing RNA-binding protein utilizing PPR motif, and use thereof

[Problem to be Solved] The object of the present invention is to develop a method of regulating a target RNA. [Solution] There is provided a fusion protein comprising a functional domain which improves the protein expression level from mRNA and a PPR protein which can bind to a target mRNA in an RNA base-selective or RNA base sequence-specific manner.

Fusion protein for improving protein expression from target mRNA

A method for designing a protein capable of binding in an RNA base selective manner or RNA base sequence specific manner is provided. The protein of the present invention is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1 (wherein Helix A is a moiety of 12-amino acid length capable of forming an &#x3b1;-helix structure, and is represented by the formula 2, wherein, in the formula 2, Ato Aindependently represent an amino acid; X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an &#x3b1;-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3, wherein, in the formula 3, the amino acids are numbered 'i' (&#x2212;1), 'ii' (&#x2212;2), and so on from the C-terminus side, provided that Lto Lmay not exist), and combination of three amino acids A, Aand Lor combination of two amino acids A, and Lis a combination corresponding to a target RNA base or base sequence.

Method for designing RNA binding protein utilizing PPR motif, and use thereof

The object of the present invention is to, by analyzing PPR proteins that act to bind to DNA with a prediction that RNA recognition rules of PPR motifs can also be used for recognition of DNA, find a PPR protein showing such a characteristic. According to the present invention, it was revealed that, with a protein that can bind in a DNA base-selective manner or a DNA base sequence-specific manner, which contains one or more, preferably 2 to 30, more preferably 5 to 25, most preferably 9 to 15, of PPR motifs having a structure of the following formula 1 (wherein, in the formula 1, Helix A is a part that can form an &#x3b1;-helix structure; X does not exist, or is a part consisting of 1 to 9 amino acids; Helix B is a part that can form an &#x3b1;-helix structure; and L is a part consisting of 2 to 7 amino acids), and having a specific combination of amino acids corresponding to a DNA base or DNA base sequence as amino acids of three positions of No. 1 A.A., No. 4 A.A., in Helix A of the formula 1 and No. 'ii' (&#x2212;2) A. A. contained in L of the formula 1, the aforementioned object could be achieved.(Helix A)-X-(Helix B)-L&#x2003;&#x2003;(Formula 1)

DNA-binding protein using PPR motif, and use thereof

A method for designing a protein capable of binding in an RNA base selective manner or RNA base sequence specific manner is provided. The protein of the present invention is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1 (wherein Helix A is a moiety of 12-amino acid length capable of forming an &#x3b1;-helix structure, and is represented by the formula 2, wherein, in the formula 2, Ato Aindependently represent an amino acid; X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an &#x3b1;-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3, wherein, in the formula 3, the amino acids are numbered 'i' (-1), 'ii' (-2), and so on from the C-terminus side, provided that Lto Lmay not exist), and combination of three amino acids A, Aand L, or combination of two amino acids A, and Lis a combination corresponding to a target RNA base or base sequence.

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Fukuoka, Japan

Yusuke Yagi