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Non-intein splicing of RB47 - Activation of an Endoribonuclease by Non-intein Protein Splicing

ABSTRACT: The poly A binding protein RB47 was purified from C. reinhardtii chloroplasts and was found to have endoribonuclease activity. MS analysis suggested that an internal non-conserved linker (NCL)sequence was absent from the native protein. Recombinant, full length RB47 protein, including the NCL did not have endoribonuclease activity while versions lacking the NCL were active. An in vitro system, utilizing recombinant RB47 and chloroplast lysates, recreated the processing of RB47 in that the NCL was excised and the flanking sequences were spliced together.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Chlamydomonas Reinhardtii

TISSUE(S): Photosynthetic Cell

SUBMITTER: Stephen Campbell

LAB HEAD: David Stern

PROVIDER: PXD004285 | Pride | 2018-10-27

REPOSITORIES: Pride

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			Action	DRS
	StephenCambell2_108C.raw	Raw
	StephenCambell2_108C_CID_semi.msf	Msf
	StephenCampbell10352147_SplicedRB47-01.msf	Msf
	StephenCampbell10352147_SplicedRB47.raw	Raw
	StephenCampbell_Band2-01.msf	Msf

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Publications

Activation of an Endoribonuclease by Non-intein Protein Splicing.

Campbell Stephen J SJ Stern David B DB

The Journal of biological chemistry 20160616 31

The Chlamydomonas reinhardtii chloroplast-localized poly(A)-binding protein RB47 is predicted to contain a non-conserved linker (NCL) sequence flanked by highly conserved N- and C-terminal sequences, based on the corresponding cDNA. RB47 was purified from chloroplasts in association with an endoribonuclease activity; however, protein sequencing failed to detect the NCL. Furthermore, while recombinant RB47 including the NCL did not display endoribonuclease activity in vitro, versions lacking the ...[more]

PMID: 27311716

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