Afp14 is involved in regulating the length of Anti‐feeding prophage (Afp). Issue 4 (16th March 2015)

• Record Type: Journal Article
• Authors: Rybakova, Daria; Schramm, Peter; Mitra, Alok K.; Hurst, Mark R. H.
• Is Part Of: Molecular microbiology. Volume 96:Issue 4(2015)
• Page Start: 815
• Page End: 826
• Publication Date: 2015-03-16
• Subjects: Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.12974
• Languages: English
• ISSNs: 0950-382X

Abstract

<abstract abstract-type="main"> <title>Summary</title> <p>The anti‐feeding prophage (Afp), a phage‐tail‐like particle that causes cessation of feeding in the New Zealand grass grub, <italic>C</italic><italic>ostelytra zealandica</italic>, is encoded by 18 open reading frames (<italic>afp1–18</italic>). C‐terminal truncations of <italic>afp14</italic> resulted in shortened Afp particles, suggesting that Afp14 is involved in Afp length determination. We constructed an Afp assembly system (<italic>afp1–18</italic>), wherein Afp14 was truncated after the N‐terminal 88 residues. This construct, when expressed <italic>in trans</italic> in <italic>E</italic><italic>scherichia coli</italic> expressing a N‐terminal 98‐amino acid Afp14 construct, yielded fully assembled Afp but no assembled Afp was detected in the case of a N‐terminal 96‐amino acid Afp14 construct. These results suggested that the 98 N‐terminal, amino acid residues of Afp14 is crucial for the initiation of Afp assembly <italic>via</italic> baseplate formation. <italic>Trans</italic>‐based expression of wild‐type <italic>afp14</italic> resulted in Afp particles of varying lengths, all of which were shorter than the wild‐type Afp particle. On the other hand, similar expression of Afp14 harboring a C‐terminal extension (KLLEH<sub>6</sub>) resulted in elongated Afp particles. This information, combined with bioinformatics data, allowed us to propose a model delineating the mechanism and role of Afp14 in the maturation of the Afp particle.</p> </abstract>