Assessing the Effects of Amino Acid Insertion and Deletion Mutations

AbstractThe Origin Recognition Complex (ORC) is an evolutionarily conserved six-subunit protein complex that binds specific sites at many locations to coordinately replicate the entire eukaryote genome. Though highly conserved in structure, ORC’s selectivity for replication origins has diverged tremendously between yeasts and humans to adapt to vastly different life cycles. In this work, we demonstrate that the selectivity determinant of ORC for DNA binding lies in a 19-amino acid insertion helix in the Orc4 subunit, which is present in yeast but absent in human. Removal of this motif from Orc4 transforms the yeast ORC, which selects origins based on base-specific binding at defined locations, into one whose selectivity is dictated by chromatin landscape and afforded with plasticity, as reported for human. Notably, the altered yeast ORC has acquired an affinity for regions near transcriptional start sites (TSSs), which the human ORC also favors.

Download Full-text

Thyrotropin Receptor Cleavage at Site 1 Involves Two Discontinuous Segments at Each End of the Unique 50-Amino Acid Insertion

Journal of Biological Chemistry ◽

10.1074/jbc.274.4.2093 ◽

1999 ◽

Vol 274 (4) ◽

pp. 2093-2096 ◽

Cited By ~ 13

Author(s):

Kunihiko Tanaka ◽

Gregorio D. Chazenbalk ◽

Sandra M. McLachlan ◽

Basil Rapoport

Keyword(s):

Amino Acid ◽

Thyrotropin Receptor ◽

Amino Acid Insertion

Download Full-text

Amino Acid Insertion Reveals a Necessary Three-Helical Intermediate in the Folding Pathway of the Colicin E7 Immunity Protein Im7

Journal of Molecular Biology ◽

10.1016/j.jmb.2009.07.085 ◽

2009 ◽

Vol 392 (4) ◽

pp. 1074-1086 ◽

Cited By ~ 9

Author(s):

Stuart E. Knowling ◽

Angelo Miguel Figueiredo ◽

Sara B.-M. Whittaker ◽

Geoffrey R. Moore ◽

Sheena E. Radford

Keyword(s):

Amino Acid ◽

Folding Pathway ◽

Immunity Protein ◽

Amino Acid Insertion ◽

Colicin E7

Download Full-text

RAISE: a simple and novel method of generating random insertion and deletion mutations

Nucleic Acids Research ◽

10.1093/nar/gnj032 ◽

2006 ◽

Vol 34 (4) ◽

pp. e30-e30 ◽

Cited By ~ 36

Author(s):

R. Fujii

Keyword(s):

Deletion Mutations ◽

Insertion And Deletion ◽

Novel Method ◽

Random Insertion

Download Full-text

Effects of DNA heterologies on bacteriophage lambda recombination.

Genetics ◽

10.1093/genetics/118.1.13 ◽

1988 ◽

Vol 118 (1) ◽

pp. 13-19

Author(s):

R K Pearson ◽

M S Fox

Keyword(s):

Bacteriophage Lambda ◽

Indirect Evidence ◽

Direct Products ◽

Base Pairs ◽

Heteroduplex Dna ◽

Deletion Mutations ◽

Genetic Crosses ◽

Insertion And Deletion ◽

Progeny Phage ◽

Substitution Mutation

Abstract Previous studies of bacteriophage lambda recombination have provided indirect evidence that substantial sequence nonhomologies, such as insertions and deletions, may be included in regions of heteroduplex DNA. However, the direct products of heterology-containing heteroduplex DNA--heterozygous progeny phage--have not been observed. We have constructed a series of small insertion and deletion mutations in the cI gene to examine the possibility that small heterologies might be accommodated in heterozygous progeny phage. Genetic crosses were carried out between lambda cI- Oam29 and lambda cI+ Pam80 under replication-restricted conditions. Recombinant O+P+ progeny were selected on mutL hosts and tested for cI heterozygosity. Heterozygous recombinants were readily observed with crosses involving insertions of 4 to 19 base pairs (bp) in the cI gene. Thus, nonhomologies of at least 19 bp can be accommodated in regions of heteroduplex DNA during lambda recombination. In contrast, when a cI insertion or deletion mutation of 26 bp was present, few of the selected recombinants were heterozygous for cI. Results using a substitution mutation, involving a 26-bp deletion with a 22-bp insertion, suggest that the low recovery of cI heterozygotes containing heterologies of 26 bp or more is due to a failure to encapsulate DNA containing heterologies of 26 bp or more into viable phage particles.

Download Full-text

Suppressor mutations in Escherichia coli methionyl-tRNA formyltransferase: Role of a 16-amino acid insertion module in initiator tRNA recognition

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.94.25.13524 ◽

1997 ◽

Vol 94 (25) ◽

pp. 13524-13529 ◽

Cited By ~ 18

Author(s):

V. Ramesh ◽

S. Gite ◽

Y. Li ◽

U. L. RajBhandary

Keyword(s):

Escherichia Coli ◽

Amino Acid ◽

Initiator Trna ◽

Trna Recognition ◽

Suppressor Mutations ◽

Amino Acid Insertion

Download Full-text

KPC-50 Confers Resistance to Ceftazidime-Avibactam Associated with Reduced Carbapenemase Activity

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00321-20 ◽

2020 ◽

Vol 64 (8) ◽

Cited By ~ 3

Author(s):

Laurent Poirel ◽

Xavier Vuillemin ◽

Mario Juhas ◽

Amandine Masseron ◽

Ursina Bechtel-Grosch ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Klebsiella Pneumoniae ◽

Clinical Isolate ◽

Sharp Reduction ◽

Content Type ◽

Amino Acid Insertion

ABSTRACT KPC-50 is a KPC-3 variant identified from a Klebsiella pneumoniae clinical isolate recovered in Switzerland in 2019. Compared to KPC-3, KPC-50 shows (i) a three-amino-acid insertion (Glu-Ala-Val) between amino acids 276 and 277, (ii) an increased affinity to ceftazidime, (iii) a decreased sensitivity to avibactam, explaining the ceftazidime-avibactam resistance, and (iv) an association with a sharp reduction of its carbapenemase activity.

Download Full-text