Methanosarcina acetivorans flap endonuclease 1 activity is inhibited by a cognate single-stranded-DNA-binding protein

Yuyen Lin, Claudia E. Guzman, Mary C. McKinney, Satish K. Nair, Taekjip Ha, Isaac K O Cann

Research output: Contribution to journalArticle

Abstract

The oligonucleotide/oligosaccharide-binding (OB) fold is central to the architecture of single-stranded-DNA-binding proteins, which are polypeptides essential for diverse cellular processes, including DNA replication, repair, and recombination. In archaea, single-stranded DNA-binding proteins composed of multiple OB folds and a zinc finger domain, in a single polypeptide, have been described. The OB folds of these proteins were more similar to their eukaryotic counterparts than to their bacterial ones. Thus, the archaeal protein is called replication protein A (RPA), as in eukaryotes. Unlike most organisms, Methanosarcina acetivorans harbors multiple functional RPA proteins, and it was our interest to determine whether the different proteins play different roles in DNA transactions. Of particular interest was lagging-strand DNA synthesis, where recently RPA has been shown to regulate the size of the 5′ region cleaved during Okazaki fragment processing. We report here that M. acetivorans RPA1 (MacRPA1), a protein composed of four OB folds in a single polypeptide, inhibits cleavage of a long flap (20 nucleotides) by M. acetivorans flap endonuclease 1 (MacFEN1). To gain a further insight into the requirement of the different regions of MacRPA1 on its inhibition of MacFEN1 endonuclease activity, N-terminal and C-terminal truncated derivatives of the protein were made and were biochemically and biophysically analyzed. Our results suggested that MacRPA1 derivatives with at least three OB folds maintained the properties required for inhibition of MacFEN1 endonuclease activity. Despite these interesting observations, further biochemical and genetic analyses are required to gain a deeper understanding of the physiological implications of our findings.

Original languageEnglish (US)
Pages (from-to)6153-6167
Number of pages15
JournalJournal of Bacteriology
Volume188
Issue number17
DOIs
StatePublished - Sep 2006
Externally publishedYes

Fingerprint

Flap Endonucleases
Methanosarcina
DNA-Binding Proteins
Oligosaccharides
Replication Protein A
Endonucleases
Peptides
Proteins
Archaeal Proteins
DNA
Archaea
Zinc Fingers
Eukaryota
DNA Replication
Oligonucleotides
DNA Repair
Genetic Recombination
Molecular Biology
Carrier Proteins
Nucleotides

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Methanosarcina acetivorans flap endonuclease 1 activity is inhibited by a cognate single-stranded-DNA-binding protein. / Lin, Yuyen; Guzman, Claudia E.; McKinney, Mary C.; Nair, Satish K.; Ha, Taekjip; Cann, Isaac K O.

In: Journal of Bacteriology, Vol. 188, No. 17, 09.2006, p. 6153-6167.

Research output: Contribution to journalArticle

Lin, Yuyen ; Guzman, Claudia E. ; McKinney, Mary C. ; Nair, Satish K. ; Ha, Taekjip ; Cann, Isaac K O. / Methanosarcina acetivorans flap endonuclease 1 activity is inhibited by a cognate single-stranded-DNA-binding protein. In: Journal of Bacteriology. 2006 ; Vol. 188, No. 17. pp. 6153-6167.
@article{6fb5d103d8c64913aa31784b75220ebd,
title = "Methanosarcina acetivorans flap endonuclease 1 activity is inhibited by a cognate single-stranded-DNA-binding protein",
abstract = "The oligonucleotide/oligosaccharide-binding (OB) fold is central to the architecture of single-stranded-DNA-binding proteins, which are polypeptides essential for diverse cellular processes, including DNA replication, repair, and recombination. In archaea, single-stranded DNA-binding proteins composed of multiple OB folds and a zinc finger domain, in a single polypeptide, have been described. The OB folds of these proteins were more similar to their eukaryotic counterparts than to their bacterial ones. Thus, the archaeal protein is called replication protein A (RPA), as in eukaryotes. Unlike most organisms, Methanosarcina acetivorans harbors multiple functional RPA proteins, and it was our interest to determine whether the different proteins play different roles in DNA transactions. Of particular interest was lagging-strand DNA synthesis, where recently RPA has been shown to regulate the size of the 5′ region cleaved during Okazaki fragment processing. We report here that M. acetivorans RPA1 (MacRPA1), a protein composed of four OB folds in a single polypeptide, inhibits cleavage of a long flap (20 nucleotides) by M. acetivorans flap endonuclease 1 (MacFEN1). To gain a further insight into the requirement of the different regions of MacRPA1 on its inhibition of MacFEN1 endonuclease activity, N-terminal and C-terminal truncated derivatives of the protein were made and were biochemically and biophysically analyzed. Our results suggested that MacRPA1 derivatives with at least three OB folds maintained the properties required for inhibition of MacFEN1 endonuclease activity. Despite these interesting observations, further biochemical and genetic analyses are required to gain a deeper understanding of the physiological implications of our findings.",
author = "Yuyen Lin and Guzman, {Claudia E.} and McKinney, {Mary C.} and Nair, {Satish K.} and Taekjip Ha and Cann, {Isaac K O}",
year = "2006",
month = "9",
doi = "10.1128/JB.00045-06",
language = "English (US)",
volume = "188",
pages = "6153--6167",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "17",

}

TY - JOUR

T1 - Methanosarcina acetivorans flap endonuclease 1 activity is inhibited by a cognate single-stranded-DNA-binding protein

AU - Lin, Yuyen

AU - Guzman, Claudia E.

AU - McKinney, Mary C.

AU - Nair, Satish K.

AU - Ha, Taekjip

AU - Cann, Isaac K O

PY - 2006/9

Y1 - 2006/9

N2 - The oligonucleotide/oligosaccharide-binding (OB) fold is central to the architecture of single-stranded-DNA-binding proteins, which are polypeptides essential for diverse cellular processes, including DNA replication, repair, and recombination. In archaea, single-stranded DNA-binding proteins composed of multiple OB folds and a zinc finger domain, in a single polypeptide, have been described. The OB folds of these proteins were more similar to their eukaryotic counterparts than to their bacterial ones. Thus, the archaeal protein is called replication protein A (RPA), as in eukaryotes. Unlike most organisms, Methanosarcina acetivorans harbors multiple functional RPA proteins, and it was our interest to determine whether the different proteins play different roles in DNA transactions. Of particular interest was lagging-strand DNA synthesis, where recently RPA has been shown to regulate the size of the 5′ region cleaved during Okazaki fragment processing. We report here that M. acetivorans RPA1 (MacRPA1), a protein composed of four OB folds in a single polypeptide, inhibits cleavage of a long flap (20 nucleotides) by M. acetivorans flap endonuclease 1 (MacFEN1). To gain a further insight into the requirement of the different regions of MacRPA1 on its inhibition of MacFEN1 endonuclease activity, N-terminal and C-terminal truncated derivatives of the protein were made and were biochemically and biophysically analyzed. Our results suggested that MacRPA1 derivatives with at least three OB folds maintained the properties required for inhibition of MacFEN1 endonuclease activity. Despite these interesting observations, further biochemical and genetic analyses are required to gain a deeper understanding of the physiological implications of our findings.

AB - The oligonucleotide/oligosaccharide-binding (OB) fold is central to the architecture of single-stranded-DNA-binding proteins, which are polypeptides essential for diverse cellular processes, including DNA replication, repair, and recombination. In archaea, single-stranded DNA-binding proteins composed of multiple OB folds and a zinc finger domain, in a single polypeptide, have been described. The OB folds of these proteins were more similar to their eukaryotic counterparts than to their bacterial ones. Thus, the archaeal protein is called replication protein A (RPA), as in eukaryotes. Unlike most organisms, Methanosarcina acetivorans harbors multiple functional RPA proteins, and it was our interest to determine whether the different proteins play different roles in DNA transactions. Of particular interest was lagging-strand DNA synthesis, where recently RPA has been shown to regulate the size of the 5′ region cleaved during Okazaki fragment processing. We report here that M. acetivorans RPA1 (MacRPA1), a protein composed of four OB folds in a single polypeptide, inhibits cleavage of a long flap (20 nucleotides) by M. acetivorans flap endonuclease 1 (MacFEN1). To gain a further insight into the requirement of the different regions of MacRPA1 on its inhibition of MacFEN1 endonuclease activity, N-terminal and C-terminal truncated derivatives of the protein were made and were biochemically and biophysically analyzed. Our results suggested that MacRPA1 derivatives with at least three OB folds maintained the properties required for inhibition of MacFEN1 endonuclease activity. Despite these interesting observations, further biochemical and genetic analyses are required to gain a deeper understanding of the physiological implications of our findings.

UR - http://www.scopus.com/inward/record.url?scp=33749022503&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749022503&partnerID=8YFLogxK

U2 - 10.1128/JB.00045-06

DO - 10.1128/JB.00045-06

M3 - Article

C2 - 16923882

AN - SCOPUS:33749022503

VL - 188

SP - 6153

EP - 6167

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 17

ER -