Characterization of a cDNA encoding a novel plant poly(A) polymerase

Jaydip Das Gupta; Qing Shun Li; A. Brian Thomson; Arthur G. Hunt

doi:10.1023/A:1006000213403

Characterization of a cDNA encoding a novel plant poly(A) polymerase

Jaydip Das Gupta, Qing Shun Li, A. Brian Thomson, Arthur G. Hunt

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

We have isolated cDNA clones encoding a novel factor (PAP-I) that is a component of a multi-subunit poly(A) polymerase from pea seedlings. The encoded protein, when isolated from appropriately engineered Escherichia coli, was active as a poly(A) polymerase, either with an associated RNA binding cofactor (PAP-III) or with free poly(A) as an RNA substrate. The latter observation indicates that PAP-I is in fact a poly(A) polymerase. PAP-I bore a striking resemblance to an as yet uncharacterized cyanobacterial protein. This observation suggested a possible chloroplast localization for PAP-I. This hypothesis was tested and found to be substantiated; immunoblot analysis identified PAP-I in chloroplast but not nuclear extracts. Our results suggest that PAP-I is a component of the machinery that adds poly(A) to chloroplast RNAs.

Original language	English
Pages (from-to)	729-734
Number of pages	6
Journal	Plant Molecular Biology
Volume	37
Issue number	4
DOIs	https://doi.org/10.1023/A:1006000213403
State	Published - Jul 1998

Bibliographical note

Funding Information:
We are grateful to Carol Von Lanken for technical assistance, and to Santanu Das Gupta and Gautam Sen for assistance in preparing antibodies. We thank Dr A.S.N. Reddy for the gift of the anti-U1 antiserum and Dr R. Houtz for anti-rbcL antibodies. This work was supported by a grant (95-37301-2041) from the USDA NRI Competitive Grant Program.

Keywords

Chloroplast RNA processing
Poly(A) polymerase

ASJC Scopus subject areas

Agronomy and Crop Science
Genetics
Plant Science

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title = "Characterization of a cDNA encoding a novel plant poly(A) polymerase",

abstract = "We have isolated cDNA clones encoding a novel factor (PAP-I) that is a component of a multi-subunit poly(A) polymerase from pea seedlings. The encoded protein, when isolated from appropriately engineered Escherichia coli, was active as a poly(A) polymerase, either with an associated RNA binding cofactor (PAP-III) or with free poly(A) as an RNA substrate. The latter observation indicates that PAP-I is in fact a poly(A) polymerase. PAP-I bore a striking resemblance to an as yet uncharacterized cyanobacterial protein. This observation suggested a possible chloroplast localization for PAP-I. This hypothesis was tested and found to be substantiated; immunoblot analysis identified PAP-I in chloroplast but not nuclear extracts. Our results suggest that PAP-I is a component of the machinery that adds poly(A) to chloroplast RNAs.",

keywords = "Chloroplast RNA processing, Poly(A) polymerase",

author = "{Das Gupta}, Jaydip and Li, {Qing Shun} and Thomson, {A. Brian} and Hunt, {Arthur G.}",

note = "Funding Information: We are grateful to Carol Von Lanken for technical assistance, and to Santanu Das Gupta and Gautam Sen for assistance in preparing antibodies. We thank Dr A.S.N. Reddy for the gift of the anti-U1 antiserum and Dr R. Houtz for anti-rbcL antibodies. This work was supported by a grant (95-37301-2041) from the USDA NRI Competitive Grant Program.",

year = "1998",

month = jul,

doi = "10.1023/A:1006000213403",

language = "English",

volume = "37",

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T1 - Characterization of a cDNA encoding a novel plant poly(A) polymerase

AU - Das Gupta, Jaydip

AU - Li, Qing Shun

AU - Thomson, A. Brian

AU - Hunt, Arthur G.

N1 - Funding Information: We are grateful to Carol Von Lanken for technical assistance, and to Santanu Das Gupta and Gautam Sen for assistance in preparing antibodies. We thank Dr A.S.N. Reddy for the gift of the anti-U1 antiserum and Dr R. Houtz for anti-rbcL antibodies. This work was supported by a grant (95-37301-2041) from the USDA NRI Competitive Grant Program.

PY - 1998/7

Y1 - 1998/7

N2 - We have isolated cDNA clones encoding a novel factor (PAP-I) that is a component of a multi-subunit poly(A) polymerase from pea seedlings. The encoded protein, when isolated from appropriately engineered Escherichia coli, was active as a poly(A) polymerase, either with an associated RNA binding cofactor (PAP-III) or with free poly(A) as an RNA substrate. The latter observation indicates that PAP-I is in fact a poly(A) polymerase. PAP-I bore a striking resemblance to an as yet uncharacterized cyanobacterial protein. This observation suggested a possible chloroplast localization for PAP-I. This hypothesis was tested and found to be substantiated; immunoblot analysis identified PAP-I in chloroplast but not nuclear extracts. Our results suggest that PAP-I is a component of the machinery that adds poly(A) to chloroplast RNAs.

AB - We have isolated cDNA clones encoding a novel factor (PAP-I) that is a component of a multi-subunit poly(A) polymerase from pea seedlings. The encoded protein, when isolated from appropriately engineered Escherichia coli, was active as a poly(A) polymerase, either with an associated RNA binding cofactor (PAP-III) or with free poly(A) as an RNA substrate. The latter observation indicates that PAP-I is in fact a poly(A) polymerase. PAP-I bore a striking resemblance to an as yet uncharacterized cyanobacterial protein. This observation suggested a possible chloroplast localization for PAP-I. This hypothesis was tested and found to be substantiated; immunoblot analysis identified PAP-I in chloroplast but not nuclear extracts. Our results suggest that PAP-I is a component of the machinery that adds poly(A) to chloroplast RNAs.

KW - Chloroplast RNA processing

KW - Poly(A) polymerase

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M3 - Article

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AN - SCOPUS:0342330401

SN - 0167-4412

VL - 37

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JO - Plant Molecular Biology

JF - Plant Molecular Biology

IS - 4

ER -

Characterization of a cDNA encoding a novel plant poly(A) polymerase

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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